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ARVO 2015 Annual Meeting Abstracts 414 Novel ophthalmic imaging Wednesday, May 06, 2015 8:30 AM–10:15 AM Exhibit Hall Poster Session Program #/Board # Range: 4079–4125/B0001–B0047 Organizing Section: Multidisciplinary Ophthalmic Imaging Group Contributing Section(s): Cornea, Retina, Visual Psychophysics/ Physiological Optics Program Number: 4079 Poster Board Number: B0001 Presentation Time: 8:30 AM–10:15 AM Relationship between visual brain connectivity and duration of blindness depends on onset of visual deprivation Kevin C. Chan1, 2, Matthew C. Murphy1, 2, Christopher Fisher2, Seong-Gi Kim1, 4, Joel S. Schuman2, 3, Amy C. Nau2. 1NeuroImaging Laboratory, University of Pittsburgh, Pittsburgh, PA; 2UPMC Eye Center, Eye and Ear Institute, Ophthalmology and Visual Science Research Center, Department of Ophthalmology, School of Medicine, University of Pittsburgh, Pittsburgh, PA; 3Department of Bioengineering, Swanson School of Engineering, University of Pittsburgh, Pittsburgh, PA; 4Center for Neuroscience Imaging Research, Institute for Basic Science, Sungkyunkwan University, Suwon, Korea (the Republic of). Purpose: Visual deprivation is known to induce plasticity of the visual system, which can be observed through alterations in functional brain connectivity (FC) by functional MRI (fMRI). How these FC changes accrue over time in congenital and acquired blindness remains uncertain. This work aimed to model the effects of prior visual experience on visual FC in blind subjects. Methods: Seven congenitally blind and 11 age-matched acquired blind subjects underwent 8 minutes of fMRI at rest using a 3 Tesla scanner. A visual FC map was constructed for each subject by computing the average correlation coefficient between the extrastriate visual cortex and each voxel in the brain. We then examined the effects of visual experience by fitting these FC maps with a linear model with predictors including duration of blindness, a dummy variable for congenital blindness, and the interaction of these two effects. T score maps of these effects were thresholded at a familywise error corrected p<0.01. Results: From the maps of voxel-wise statistical testing (Fig. 1), more than 6 times as many voxels in the brain show a significant group by duration interaction effect (row 4) compared to a simple correlation with duration of blindness (row 2), indicating that the relationship between visual FC and duration of blindness is significantly different between congenital and acquired blindness. In general, the sign of the correlation between FC and duration of blindness in congenital subjects is opposite to that in acquired subjects. Similar findings are observed when using striate cortex instead of extrastriate cortex as the seed region of interest (data not shown). Conclusions: This work represents an early step toward understanding plasticity in the visual system and how it depends upon prior visual experience. These results suggest that alterations in FC due to visual deprivation progress over time but in opposite directions between congenital and acquired blindness. Our findings indicate that longitudinal measures of FC and not only FC alone may be essential for characterizing the state of the visual system. Figure 1. Summary of voxel-wise statistical testing (FWE corrected p<0.01). Row 1: T score for average visual FC map for all subjects. Row 2: T score for significant relationship between FC and duration of blindness. Row 3: T score for group-wise differences. Row 4: T score for group by duration interaction. Commercial Relationships: Kevin C. Chan, None; Matthew C. Murphy, None; Christopher Fisher, None; Seong-Gi Kim, None; Joel S. Schuman, Zeiss, Inc. (P); Amy C. Nau, None Support: NIH Grant P30 EY008098 and T32-EY017271 (Bethesda, MD); US Department of Defense DM090217; Alcon Research Institute Young Investigator Grant (Basel, Switzerland); Eye and Ear Foundation (Pittsburgh, PA); Research to Prevent Blindness (New York, NY); Aging Institute Pilot Seed Grant, University of Pittsburgh; Louis J. Fox Center for Vision Restoration of UPMC and the University of Pittsburgh Program Number: 4080 Poster Board Number: B0002 Presentation Time: 8:30 AM–10:15 AM In vivo MR Diffusion Weighted Imaging at 7 Tesla – feasibility study in healthy subjects and patients with different ophthalmic disease Tobias Lindner1, Katharina Paul2, Andreas Graessl2, Jan Rieger3, Till Huelnhagen2, Paul-Christian Krueger4, Anselm Juenemann5, Soenke Langner4, Thoralf Niendorf2, 3, Oliver Stachs5. 1Preclinical Imaging Research Group, University Medicine Rostock, Rostock, Germany; 2 Berlin Ultrahigh Field Facility, Max-Delbrueck Center for Molecular Medicine, Berlin, Germany; 3MRI Tools, Berlin, Germany; 4Institute for Diagnostic Radiology and Neuroradiology, University Medicine Greifswald, Greifswald, Germany; 5Department of Ophthalmology, University Medicine Rostock, Rostock, Germany. Purpose: Diffusion-weighted imaging (DWI) of the eye and orbit is an emerging MRI application for ophthalmic research, preclinical imaging and diagnostic radiology. DWI probes and depicts selfdiffusion of water in tissue on a microscopic scale and holds the promise to enhance diagnostic accuracy and provide further physiological information for different ophthalmic disease. The purpose of this study was to show the feasibility of in vivo DWI at 7 T with diffusion sensitized multi-shot RARE sequences. Methods: In vivo MR imaging and DWI was performed at 7T in healthy subjects (n=5), patients with uveal melanoma and/or retinal detachment (n=5) and a patient with optic nerve glioma. A dedicated six element transmit and receive coil array was employed at 7.0 T and high resolution T1 weighted 3D FLASH imaging (TR=710.3ms, ©2015, Copyright by the Association for Research in Vision and Ophthalmology, Inc., all rights reserved. Go to iovs.org to access the version of record. For permission to reproduce any abstract, contact the ARVO Office at [email protected]. ARVO 2015 Annual Meeting Abstracts TE=3.6ms, spatial resolution = (0.3x0.3x1.0)mm3 and T2-weighted RARE imaging (TR=2940ms, TE=54ms, spatial resolution = (0.25x0.28x1.0)mm3 were conducted for anatomical imaging. For ADC mapping diffusion sensitized multi-shot RARE was used (7.0 T: TR=3000ms, TE=133ms, spatial resolution = (0.4x0.4x3.0)mm3). For diffusion sensitization b-values ranging from b=0 s/mm2 to b=400 s/ mm2 were employed. To reduce eye movement during imaging the subjects followed an optoacoustic trigger protocol. Results: The in vivo ADC map of patients with ocular mass clearly delineates hypointense tumor from the surrounding modest hypointense hemorrhage and the hyperintense vitreous body. These ADC differences induced an ample contrast between the subretinal hemorrhage and the tumor. ADC map of orbita in a patient with optic nerve glioma showed ample ADC contrast versus the subarachnoid space. The ADC map of the eye and optic nerve yielded a delineation of the tumor which is superior to the contrast in the T2 weighted images. Conclusions: This work demonstrates the feasibility of in vivo MR microscopy and diffusion sensitized ms-RARE of the eye and optic nerve at 7.0T including T1w, T2w anatomical imaging along with DWI and ADC mapping. This study showed that ms-RARE based techniques offer immunity to B0 inhomogeneities and hence are particularly suited for ophthalmic DWI. This is of clinical relevance since single-shot echo planar imaging is prone to magnetic susceptibility artifacts induced by the air filled nasal cavities and frontal sinuses surrounding the eye. Commercial Relationships: Tobias Lindner, None; Katharina Paul, None; Andreas Graessl, None; Jan Rieger, MRI Tools (E); Till Huelnhagen, None; Paul-Christian Krueger, None; Anselm Juenemann, None; Soenke Langner, None; Thoralf Niendorf, MRI Tools (S); Oliver Stachs, None Program Number: 4081 Poster Board Number: B0003 Presentation Time: 8:30 AM–10:15 AM Fundus Changes in Highly Myopic Eyes with Different Shapes Identified by High-resolution Three-dimensional Magnetic Resonance Imaging Xinxing Guo1, Ou Xiao1, Yanxian Chen1, Mingguang He1, 2. 1Division of Preventive Ophthal, Zhongshan Ophthalmic Center, Guangzhou, China; 2Centre for Eye Research Australia, University of Melbourne, Melbourne, VIC, Australia. Purpose: High myopia is often associated with elongation and distortion of the globe, and at greater risk of developing pathologic changes. Lack of topographic and morphologic assessments hinders the understanding towards the natural history and mechanism in its development. We evaluated the fundus changes in high myopia by analyzing the eye shapes using three-dimensional (3D) magnetic resonance imaging (MRI) in a case series study. Methods: A total of 95 patients with high myopia (spherical power ≤ -6.00D) in both eyes underwent a series of ophthalmic examinations. Eye shapes were categorized into spherical, ellipsoidal, cylindrical, nasal-distorted, temporal-distorted and barrel-shaped according to the inferior view from T2-weighted 3D MRI images (Achieva 3.0T, Philips Medical Systems, Best, the Netherlands); posterior staphyloma was also identified. Fundus lesions were determined using two 45° photos centered at macula and optic nerve by dilated fundus photography (Canon Inc., Tokyo, Japan). The distributions of various fundus characteristics in different eye shape categories were analyzed using one-way ANOVA and Mann-Whitney Wilcoxon tests. Results: The participants consisted 48.4% females, had a mean age of 32.0±14.0 years old, spherical equivalence of -11.00±7.44D and axial length (AL) of 28.18±1.73mm. The same ocular shapes between the two eyes were observed in 72.6% of the patients. The most predominant shape in the right eye was spherical (54.3%), followed by nasal-distorted (16.0%) and cylindrical (14.9%). Barrel-shaped eyes had the longest AL and most outstanding fundus changes. Diffuse chorioretinal atrophy and larger peripapillary atrophy (PPA) were more frequently observed in barrel-shaped (100.0%, 4.7±2.7 disc area, DA) and nasally distorted (73.3%, 2.5±2.5 DA) eyes; While diffuse (77.3%) and patchy (22.7%) chorioretinal atrophy, as well as fundus staphyloma (72.7%) were more common in eyes with posterior staphyloma (n=22) determined by 3D MRI. Conclusions: Barrel-shaped eyes present most significant visionthreatening conditions, while eyes with posterior staphyloma display more severe chorioretinal atrophy. Our findings suggest different topographic and morphologic patterns may be involved in the development of high myopia. Different eye shapes and corresponding fundus images Commercial Relationships: Xinxing Guo, None; Ou Xiao, None; Yanxian Chen, None; Mingguang He, None ©2015, Copyright by the Association for Research in Vision and Ophthalmology, Inc., all rights reserved. Go to iovs.org to access the version of record. For permission to reproduce any abstract, contact the ARVO Office at [email protected]. ARVO 2015 Annual Meeting Abstracts Program Number: 4082 Poster Board Number: B0004 Presentation Time: 8:30 AM–10:15 AM Eye morphology quantitated by magnetic resonance imaging in C57Bl/6 mice Tomasz J. Choragiewicz1, Michal Fiedorowicz2, Marlena WelniakKaminska2, Jaroslaw Orzel2, 3, Piotr Bogorodzki2, 3, Robert Rejdak1, 2 , Pawel Grieb2. 1Department of General Ophthalmology, Medical University of Lublin, Lublin, Poland; 2Mossakowski Medical Research Centre, Polish Academy of Sciences, Warsaw, Poland; 3 Faculty of Electronics and Information Technology, Warsaw University of Technology, Warsaw, Poland. Purpose: Evaluating eye dimensions is crucial in studies of animal models of eye diseases. However, performing these measurements in alive mice is challenging. The aim of this study was to develop a method for quantitating eye morphology using planar surface MRI coil. Methods: Aged (18 months) C57Bl/6 female mice (n=5) were anaesthetized with isoflurane (4% in oxygen – induction, 1.52% - maintenance) and placed in 7T small animal-dedicated magnetic resonance tomograph (BioSpec 70/30USR; Bruker BioSpin, Ettlingen, Germany). Small planar surface receive coil (internal diameter=10 mm, Bruker BioSpin, Ettlingen, Germany) was placed over left eye of the imaged animal. High resolution structural imaging with TURBORARE T2 sequence (TR=2700ms, TEeff=30ms, RARE factor=8, NA=12, spatial resolution=0.062mm x 0.062mm, slice thickness=0.3mm, slices=7, scan time=16min) was performed. Basic ocular dimensions and anterior chamber angle were measured manually using OsiriX software (Pixmeo, SARL, Bernex, Switzerland). Results: Mean anterior chamber depth in C57Bl/6 mice was 0.464 mm (SD ±0.040) and anterior chamber angle (ACA) 24.98° (SD ±5.599), vitreous chamber depth (VCD) 0.465 mm (SD ±0.033), axial length (AL) 3.586 mm (SD ±0.079) and horizontal length (HL) 3.394 mm (SD ±0.061) of the eyeball, lens thickness (LT) 2.146 mm (SD ±0.059) and optic nerve diameter (ON) 0.250 mm (SD ±0.0520), retinal thickness (RT) 0.206 mm (SD ±0.018). Conclusions: High resolution magnetic resonance imaging of the eye with use of small planar coil provide reproducible and consistent measures of key dimensions of eyeball. These results are consistent with results obtained with optical coherence tomography in C57Bl/6 mice [Chou et al. 2011, IOVS 52:3604-3612]. Commercial Relationships: Tomasz J. Choragiewicz, None; Michal Fiedorowicz, None; Marlena Welniak-Kaminska, None; Jaroslaw Orzel, None; Piotr Bogorodzki, None; Robert Rejdak, None; Pawel Grieb, None Support: The study was supported by Polish National Science Centre grant No. 2012/07/D/NZ4/04199, Tomasz Choragiewicz was supported by Allergan European Retina Panel II. Program Number: 4083 Poster Board Number: B0005 Presentation Time: 8:30 AM–10:15 AM Through the Eyes of the Retinal Surgeon: Google Glass for Recording Scleral Buckling Surgery Michelle E. Wilson, Ehsan Rahimy, Sunir J. Garg. Ophthalmology, Wills Eye Hospital, Philadelphia, PA. Purpose: Google Glass (Google Inc., Mountain View, CA), with its point-of-view (POV)-style recording, has the potential to revolutionize the field of medical education, allowing trainees to see the various steps of surgical procedures directly from viewpoint of the operating surgeon. Unlike intraocular surgery, which benefits from the video documenting capabilities of modern ophthalmic surgical microscopes, scleral buckling surgery has traditionally been difficult to record, limiting access to a valuable educational modality. We report our experience using the Google Glass to record scleral buckling surgery. Methods: We recorded and subsequently reviewed both still images and videos of 3 primary scleral buckling surgeries. We qualitatively assessed the ability of the recorded material to document key steps in the scleral buckling procedure, noting overall image quality as well as factors limiting image quality. Results: Google Glass was effective in recording the operative field during each step of the scleral buckling procedure. Still images (Figure 1) and video clips (Video, Supplemental) obtained provided sufficient detail to demonstrate the key steps of the procedure. Optimal image quality was limited by several factors including overor under- illumination and lack of magnification. Conclusions: The field of medical education has benefited from new technologies that allow trainees to visualize complex surgical procedures from the viewpoint of the primary surgeon. The recent introduction of Google Glass extends this capability to surgical procedures that do not require an operating microscope and have traditionally been difficult to record for the purposes of medical education. Although limited by lighting and magnification, the point of view recordings created using Google Glass during retinal buckling surgery are able to document the key steps of the scleral buckling procedure, providing a valuable tool for surgical teaching. Scleral buckling surgery as recorded by Google Glass Commercial Relationships: Michelle E. Wilson, None; Ehsan Rahimy, None; Sunir J. Garg, None Program Number: 4084 Poster Board Number: B0006 Presentation Time: 8:30 AM–10:15 AM Real-Time Swept-Source Microscope-Integrated versus HandHeld Spectral-Domain Optical Coherence Tomography during Macular Hole Surgery Michael I. Seider, Oscar Carrasco-Zevallos, Brenton Keller, Joseph A. Izatt, Cynthia A. Toth, Christian Viehland. Duke University, Durham, NC. Purpose: To compare the clinical utility of a novel real-time swept-source microscope-integrated optical coherence tomography (SS-MIOCT) system to hand-held spectral-domain OCT (HH-OCT) during macular hole repair. Methods: All patients undergoing macular hole repair from 9/1/2014 – 10/31/2014 at the Duke Eye Center by a senior Vitreoretinal Surgeon (CT) who gave consent were included. During macular surgery, SS-MIOCT was used for all patients and allowed realtime imaging through the operating microscope. The system has ©2015, Copyright by the Association for Research in Vision and Ophthalmology, Inc., all rights reserved. Go to iovs.org to access the version of record. For permission to reproduce any abstract, contact the ARVO Office at [email protected]. ARVO 2015 Annual Meeting Abstracts acquisition and processing rates of 2-10 volumes per second and permitted the creation of volumetric renderings of surgical maneuvers over time. HH-OCT (Bioptigen Inc., Morrisville, NC, USA) was also performed in the operating room at pauses in surgery immediately before and at the completion of macular surgery. The images produced by both systems were compared post-operatively. Results: Five female and 1 male patient with average age 70 years were included, contributing three right and three left eyes in total. SS-MIOCT was able to obtain high-quality, volumetric renderings of surgical maneuvers in real-time without causing an interruption in surgery. Particularly striking images documented instruments brushing the retinal surface and forceps peeling internal limiting membrane (ILM). SS-MIOCT scans allowed for the characterization of anatomy-instrument relationships such as the distance of an instrument from the retinal surface, the retinal deformation created during scraping and the characteristics of elevated membranes volumetrically and over time. Hand-held OCT produced high-resolution two-dimensional scans at pauses during surgery. These images allowed for the accurate characterization of the morphology of the macular holes before and after ILM peeling, and the confirmation of removal of epiretinal membranes. For the cases of ILM autografting, HH-OCT created useful images confirming the location of the autograft over the macular hole. Conclusions: SS-MIOCT appears to have significant advantages over HH-OCT during macular hole surgery. SS-MIOCT was able to capture volumetric scans of surgical maneuvers in real-time without interrupting surgery whereas HH-OCT was useful for obtaining highquality static images of retinal anatomy during breaks in surgery. Commercial Relationships: Michael I. Seider, None; Oscar Carrasco-Zevallos, None; Brenton Keller, None; Joseph A. Izatt, None; Cynthia A. Toth, None; Christian Viehland, None Program Number: 4085 Poster Board Number: B0007 Presentation Time: 8:30 AM–10:15 AM Real-time 4D Stereoscopic Visualization of Human Ophthalmic Surgery with Swept-Source Microscope Integrated Optical Coherence Tomography Oscar Carrasco-Zevallos1, Brenton Keller1, Christian Viehland1, Liangbo Shen1, Gar Waterman1, Crystal Chukwurah1, Paul Hahn2, Anthony N. Kuo2, Cynthia A. Toth2, 1, Joseph A. Izatt1, 2. 1Biomedical Engineering, Duke University, Durham, NC; 2Opthalmology, Duke University Medical Center, Durham, NC. Purpose: Ophthalmic surgery is performed with a microscope that provides limited depth perception. Surgeons often rely on indirect cues for depth information. Current intraoperative Spectral-Domain OCT systems are limited to cross-sectional real-time imaging. Ophthalmic surgery is performed in a 3D surgical field; therefore, a real-time 3D micron-scale imaging modality could be useful for surgical feedback and guidance. We report on the development of 4D (volumetric imaging + time) Microscope Integrated OCT (4D MIOCT) for real-time volumetric visualization of human ophthalmic surgery. Methods: The MIOCT sample arm enabled concurrent OCT and operating microscope imaging. The system employed a custom swept-source OCT engine operating at 1060 nm. GPU-based custom software enabled real-time acquisition, processing, and rendering of volumetric images at 100k A-lines/second. Volumetric frame rates varied between 2-10 Hz. A custom microscope integrated stereoscopic heads-up display (HUD) allowed for visualization of MIOCT volumes through the surgical binoculars. 4D MIOCT imaging was performed in 25 human surgeries (7 anterior segment, 18 vitreoretinal). Vitreoretinal cases imaged included macular hole, retinal detachment, and epi-retinal membrane (ERM) procedures. Anterior segment cases imaged included cataract, deep anterior lamellar keratoplasty, and Descemet stripping endothelial keratoplasty procedures. Results: Figure 1 shows 4D MIOCT imaging of an ERM peel with surgical forceps during a macular hole surgical case. The time stamps for each volume in the time series are in seconds. B-scans are shown below each corresponding volume. The location of the B-scans is denoted by the white rectangle on the volume view. Surgical camera frames are shown as well with a yellow rectangle denoting the MIOCT field of view. Figure 2 illustrates 4D MIOCT imaging of a cataract case. A-B depict volumes and B-scans acquired at different stages of cataract surgery. (A) shows the intact cataract. (B) shows cataract fragmentation with a phacoemulsification needle (red arrow). (C) shows the anterior segment after cataract removal. (D) shows intraocular lens (green arrow) insertion. Conclusions: Real-time, volumetric, micron-scale visualization of human ophthalmic surgery was performed with 4D MIOCT. 4D MIOCT imaging of an ERM peel. 4D MIOCT imaging of cataract surgery. Commercial Relationships: Oscar Carrasco-Zevallos, None; Brenton Keller, None; Christian Viehland, None; Liangbo Shen, None; Gar Waterman, None; Crystal Chukwurah, None; Paul Hahn, None; Anthony N. Kuo, Bioptigen (P); Cynthia A. Toth, Alcon (P), Bioptigen (F), Duke University (P), Genetech (F); Joseph A. Izatt, Bioptigen (I), Bioptigen (P), Bioptigen (S) Support: NIH EY023039 Clinical Trial: NCT01588041 ©2015, Copyright by the Association for Research in Vision and Ophthalmology, Inc., all rights reserved. Go to iovs.org to access the version of record. For permission to reproduce any abstract, contact the ARVO Office at [email protected]. ARVO 2015 Annual Meeting Abstracts Program Number: 4086 Poster Board Number: B0008 Presentation Time: 8:30 AM–10:15 AM Impact of Microscope Integrated OCT on ophthalmology resident performance of anterior segment maneuvers in model eyes Bozho -. Todorich1, Christine Shieh1, Philip DeSouza1, Oscar Carrasco-Zevallos2, David Cunefare1, Joseph A. Izatt2, Sina Farsiu1, Prithvi Mruthyunjaya1, Anthony N. Kuo1, Cynthia A. Toth1, 2. 1 Ophthalmology, Duke University, Durham, NC; 2Bioengineering, Duke University, Durham, NC. Purpose: The integration of swept-source optical coherence tomography (SS OCT) in the operating microscope enables realtime, tissue-level imaging to aid in ophthalmic microsurgery. In this prospective randomized controlled study, we evaluated the impact of SS microscope-integrated OCT technology (MI OCT) on ophthalmology residents’ performance in anterior segment surgery. Methods: Ophthalmology residents (N=14) were recruited, stratified by year of training, and randomized to perform the following four anterior segment maneuvers on porcine eyes without (Group A) or with (Group B) direct MI OCT guidance: 50% and 90% depth corneal suture passes, repair of vertical linear corneal laceration via suturing at 90% thickness, and creation of a tri-planar clear corneal incision. Both groups of residents then repeated the maneuvers without MI OCT guidance. Lastly, the non MI-OCT controls (Group A) repeated all maneuvers under direct MI OCT guidance. Volumetric OCT scans acquired at the end of each maneuver were manually segmented to compute point of maximal depth of each corneal pass and tri-planar corneal incision profile. Differences between Group A and B were compared using one-way ANOVA (Figure 1A). Subjective feedback through a survey was also obtained from each resident after they had completed all maneuvers (Figure 1B). Results: Figure 1 shows the results from Group A (no MIOCT guidance) and Group B (MIOCT guidance). Residents operating with direct MI OCT feedback demonstrated enhanced performance in depth-based anterior segment maneuvers compared to the control group (p<0.0001). Residents trained with MI OCT continued to outperform the controls when operating without direct MI OCT feedback (p<0.0001) (Fig 1A). Surgical performance of residents who were trained and tested without MI OCT improved to equivalent performance when given MI OCT feedback. Resident surgeons rated subjective experience of using MI OCT very favorably and on average are “more likely” to use it in their future practice (Fig 1B). Conclusions: Real-time MI OCT guidance improved depth based surgical maneuvers. MI OCT may be a useful adjunct in surgical education of ophthalmology residents. Figure 1. Surgical performance (A) and subjective feedback (B) of ophthalmology residents with and without MI OCT feedback. Commercial Relationships: Bozho -. Todorich, None; Christine Shieh, None; Philip DeSouza, None; Oscar Carrasco-Zevallos, None; David Cunefare, None; Joseph A. Izatt, Bioptigen (I), Bioptigen (P), Bioptigen (S); Sina Farsiu, None; Prithvi Mruthyunjaya, None; Anthony N. Kuo, Bioptigen (P); Cynthia A. Toth, Alcon (P), Bioptigen (F), Genentech (F) Support: R01-EY02039 (Toth/Izatt) and Research to Prevent Blindness grant (DeSouza) Program Number: 4087 Poster Board Number: B0009 Presentation Time: 8:30 AM–10:15 AM Macroarchitectural Intraoperative Retinal Alterations Following Membrane Peeling Visualized with Intraoperative OCT in the PIONEER Study Jaehong Han1, 2, Sunil K. Srivastava2, Peter K. Kaiser2, Rishi P. Singh2, Justis P. Ehlers2. 1Case Western Reserve University School of Medicine, Cleveland Heights, OH; 2Ophthalmic Imaging Center, Cole Eye Institute of Cleveland Clinic, Cleveland, OH. Purpose: To assess the retinal macroarchitectural alterations that occur during membrane peeling for vitreomacular interface (VMI) disorders utilizing intraoperative OCT (iOCT) Methods: The PIONEER study is a prospective study examining iOCT in ophthalmic surgery. Eyes undergoing membrane peeling were evaluated with video/iOCT correlation. Clinical variables assessed included membrane peeling technique (e.g., diamond-dusted membrane scraper [DDMS], forceps). iOCT images were reviewed for macroarchitectural changes (e.g., inner and full-thickness retinal changes). A microscope mounted portable SD-OCT probe (Bioptigen, Research Triangle Park, NC) was used to obtain preincision and post-peel iOCT images. Postoperative OCT images were analyzed at 1 week and 1 month. Results: Of the total 163 eyes, post-peel retinal changes were noted by iOCT in 45 eyes (28%). Inner retinal elevations were noted in 38 eyes (23%), and 8 eyes (5%) had full-thickness retinal elevations. Based on video/iOCT correlation, 95% of the retinal changes correlated with instrument utilization. Direct peel initiation resulted in 47% of the changes and indirect pulling during completion of peel resulted in 48% of the changes. The retinal elevations occurred in 33% of the eyes when using forceps-only and in 23% of the eyes when using both forceps and a DDMS (p= 0.16, chi square). At 1-week post-op, 19% of the retinal changes persisted as focal inner retinal thickening or thinning. No full-thickness retinal changes ©2015, Copyright by the Association for Research in Vision and Ophthalmology, Inc., all rights reserved. Go to iovs.org to access the version of record. For permission to reproduce any abstract, contact the ARVO Office at [email protected]. ARVO 2015 Annual Meeting Abstracts persisted. At 1-month post-op, 60% of inner retinal thickening had resolved, while all focal retinal thinning persisted. Conclusions: Significant subclinical macroarchitectural changes occur during membrane peeling for VMI conditions that are directly related to instrument manipulations. iOCT technology is able to identify these changes. There was a trend towards increased alterations with forceps-only technique compared to using a DDMS for peel initiation. Inner retinal changes persisted in many eyes at 1 month postoperatively. Further research is needed on the visual effects of these changes and how to minimize these changes during surgery. A: preincision iOCT image; B: post-peel iOCT image; C: iOCTgenerated fundus map; D: correlated surgery video Program Number: 4088 Poster Board Number: B0010 Presentation Time: 8:30 AM–10:15 AM Evaluation of a Surgical Microscope Interfaced SD-OCT system for Anterior Segment Surgery Marco Ruggeri1, Florence Cabot1, Sonia H. Yoo1, Jean-Marie A. Parel1, 2. 1Ophthalmology, Bascom Palmer Eye Inst, Univ of Miami, Miami, FL; 2Vision Cooperative Research Centre, Brien Holden Vision Institute, UNSW, Sydney, NSW, Australia. Purpose: To evaluate the use of an intraoperative SD-OCT system that attaches to an ophthalmic operating microscope (OPMI) for ondemand OCT guided anterior segment (AS) surgery. Methods: We used a commercial prototype (Bioptigen, Inc.) that consists of a compact OCT delivery console connected to a transportable cart that houses an SD-OCT engine, a computer for image acquisition and processing and a monitor for image display. The system provides near 9mm axial resolution over an imaging depth of 15.4 mm in air. The scanning console can attach to several commercial OPMI models and includes a proprietary microscope objective with a focal length of 175 mm, a two axes OCT scanning mechanism and the optics to combine the OCT scanning beam with the illumination and viewing paths of the host OPMI. The delivery console is setup by removing the original microscope objective lens from the host OPMI and by securing the scanning head enclosure to the dovetail connection of the host OPMI. Prior to surgery, the OCT delivery unit was attached to the OPMI and the cart was deployed near the bedside so that intraoperative OCT images of the surgical site could be displayed in real-time to the surgeon using the computer monitor. Results: Under an IRB approved protocol, 4 patients who underwent AS surgical procedures including cataract and DSAEK surgeries were imaged intraoperatively. Surgeries were uneventful. The system enabled displaying the images to the surgeon without interrupting the surgical maneuvers or blocking the OPMI view. The distance of 167 mm between the microscope objective and the patient eye provided enough room for the surgeon to operate comfortably. The compact surgical OCT interface did not interfere with the sterile field and the standard operations performed by the surgical staff. Preoperatively, the system enabled to confirm that the AS did not show any abnormality. Throughout DSAEK surgeries, the long imaging depth and the volumetric capability of the SD-OCT system enabled to verify optimal adhesion of the entire graft to the host cornea in 3D. Throughout cataract surgery the system enabled to verify the integrity of the capsule and the final position of the intraocular lens. Conclusions: The intraoperative SD-OCT system can be easily and safely used to assist the surgeon for AS surgeries. This tool has potential to improve the outcome of DSAEK and help with challenging cases of cataract surgery. A, B: preoperative OCT-generated fundus map and OCT image; C, D: postoperative OCT-generated fundus map and OCT image Commercial Relationships: Jaehong Han, None; Sunil K. Srivastava, Allergan (R), Baucsh and Lomb (C), Baucsh and Lomb (R), Bioptigen (P), Carl Zeiss Meditec (C), Leica (C), Synergetics (P); Peter K. Kaiser, Alcon (C), Bausch and Lomb (C), Bayer (C), Carl Zeiss Meditec (C), Genentech (C), Novartis (C), Ophthotech (C), Oraya (C), Regeneron (C), Topcon (C); Rishi P. Singh, Carl Zeiss Meditec (C); Justis P. Ehlers, Alcon (C), Bioptigen (C), Bioptigen (F), Bioptigen (P), Genentech (F), Leica (C), Leica (F), Synergetics (P), Thrombogenics (C), Thrombogenics (F), Zeiss (C), Zeiss (F) Support: NIH/NEI K23-EY022947-01A1 (JPE); Ohio Department of Development TECH-13-059 (JPE); Thrombogenics research grant: THROM1403JE ©2015, Copyright by the Association for Research in Vision and Ophthalmology, Inc., all rights reserved. Go to iovs.org to access the version of record. For permission to reproduce any abstract, contact the ARVO Office at [email protected]. ARVO 2015 Annual Meeting Abstracts Conclusions: Integrating automated lateral and depth tracking with iOCT will allow real-time imaging of surgical dynamics. Improved understanding of tissue-instrument interactions during conventional ophthalmic surgical maneuvers may elucidate mechanisms of tissue repair, be predictive of clinical outcomes, and lead to novel iOCTguided surgical techniques. Commercial Relationships: Marco Ruggeri, Bioptigen Inc. (F); Florence Cabot, None; Sonia H. Yoo, None; Jean-Marie A. Parel, Bioptigen Inc. (F) Support: Bioptigen Inc; NIH P30EY14801 (Center Grant); Florida Lions Eye Bank; Drs KR Olsen and ME Hildebrandt, Research to Prevent Blindness; The Henri and Flore Lesieur Foundation (JMP). Program Number: 4089 Poster Board Number: B0011 Presentation Time: 8:30 AM–10:15 AM Real-time dynamic depth tracking for arbitrarily long range OCT imaging and surgical instrument tracking using a Fourier domain optical delay line Mohamed El-Haddad, Yuankai Tao. Ophthalmic Research, Cleveland Clinic Foundation, Shaker Heights, OH. Purpose: Intraoperative optical coherence tomography (iOCT) allows visualization of tissue microstructure, and provides real-time feedback for clinical decision making during ophthalmic surgery. However, the potential for iOCT-guided surgical maneuvers has been limited both by imaging speed, which can only display single cross-sectional images at video-rates, and imaging range, which is on the order of 2 mm for SDOCT and 8 mm for SSOCT. Last year, we presented novel lateral tracking technology that automatically centered the iOCT scan field on the tip of ophthalmic surgical instruments. Here, we demonstrate a digitally controlled optical delay that allows real-time depth tracking. Integration of automated lateral and depth tracking with iOCT will allow for dynamic threedimensional field-of-view imaging of ophthalmic surgical dynamics. Methods: A reference arm based on a Fourier domain optical delay line (FDODL), was designed to provide a digitally controlled optical delay for iOCT. A free-space optical delay on a motorized stage was added to the FDODL path to further extend the available tracking range. A control algorithm handles the actuation hand-off between the stage and the FDODL to allow for fast scanning over a short range, and slower scanning over a long range (Fig. 1). The FDODL allowed inter-line depth tracking over >16 mm depth range with a 100 ms small-angle step response (Fig. 1, δz) while the motorized stage allowed for inter-frame scanning over a 100 mm range at speeds up to 100 mm/s (Fig. 1, Δz). Results: As a demonstration, we imaged a metal surface placed at ~62 deg. to the horizontal (Fig. 1). Without tracking, small part of the surface was visible (Fig. 1b, blue). The FDODL was then scanned by a saw-tooth function, effectively compensating for the tilt (Fig. 1b, red). The A-Scans in the latter represent different depths, in linear proportion to the applied voltage (3.7 mm/volt), which was corrected for in post-processing to reflect the true image over the entire range of the FDODL. Commercial Relationships: Mohamed El-Haddad, None; Yuankai Tao, None Program Number: 4090 Poster Board Number: B0012 Presentation Time: 8:30 AM–10:15 AM Utility of the intraoperative OCT during posterior segment surgery Michele Coppola, Federico Solignani. Ophthalmology, Azienda Ospedaliera di Desio e Vimercate (MB), Milano, Italy. Purpose: Intraoperative OCT is a new surgical tool helping surgeons, improving intraoperative safety and allowing a real-time feedback from all ocular structures: epiretinal membranes, internal limiting membrane, outer retinal layers. Our purpose is showing, directly through videos, the easy use of iOCT and its add-on, like checking the closure of macular holes during surgery ( high myopic macular hole closure by external indentation), the complete removal of ERM and ILM even without using dyes twice. Methods: ZEISS RESCAN Lumera 700 (Carl Zeiss Meditec, Dublin, Calif., USA) - with integrated intra-operative OCT technology and Callisto eye software (Carl Zeiss Meditec, Dublin, Calif., USA) can be used during all surgical procedures performed on anterior segment, glaucoma, and retina. The wavelength of spectral domain iOCT is 840 nm, the scanning speed is 27000 A-scans per second with an A-scan depth of 2.0 mm, an axial resolution of 5.5 μm in tissue. Scan length is adjustable from 3 to 16 mm, and scan rotation ©2015, Copyright by the Association for Research in Vision and Ophthalmology, Inc., all rights reserved. Go to iovs.org to access the version of record. For permission to reproduce any abstract, contact the ARVO Office at [email protected]. ARVO 2015 Annual Meeting Abstracts is adjustable over 360°. Image acquisition is possible by recording movies or taking pictures. Results: The new machinery, iOCT, improves the safety and the surgeon’ self confidence during every surgical procedure. Starting new era that opens other surgical scenarios: surgeons can work with anatomical guide. iOCT shows anatomical reperies of the eye directly in surgical field overlaying the main image. It is controlled by the foot switch, allowing the surgeon to move indipendently from other members of the surgical staff thus enhancing the theatre safety. Conclusions: In conclusion, the importance of our video is related to the use of a new technique that allows the use of OCT during surgery and in selected cases of diagnosis as well. Further studies are certainly necessary for a better comprehension of uses and limitations of iOCT, but our videos for the first time demonstrate possible new applications. iOCT showing epiretinal membrane interrruption and its retraction after sharp needle scraping. Commercial Relationships: Michele Coppola, None; Federico Solignani, None Program Number: 4091 Poster Board Number: B0013 Presentation Time: 8:30 AM–10:15 AM Intraoperative Optical Coherence Tomography (iOCT) In Newborn And Infant Children With Anterior Segment Anomalies Sebastian E. Siebelmann, Philipp Steven, Manuel Hermann, Thomas Dietlein, Claus Cursiefen. Department of Ophthalmology, University of Cologne, Cologne, Germany. Purpose: Examination of newborn or infant children with anterior segment anomalies is often challenging due to limited compliance, so that an examination under general anesthesia is preferred. However, intraoperative imaging tools are so far limited. Therefore we evaluated intraoperative Optical Coherence Tomography (iOCT) as a tool for intraoperative imaging, diagnostics and surgery monitoring in newborn and infants with anterior segment anomalies and corneal opacities. Methods: Retrospective case series of four newborn and infants with anterior segment anomalies (2 children with congenital corneal opacities, 1 child with congenital cataract and one with iridocorneal synechiae), who underwent examinations under general anesthesia, using a commercially available 840nm OCT, mounted to an operating microscope (iOCT; OptoMedical Technologies, Luebeck, Germany). Mean age was 6 years (range: 6 month – 14 years). Results: Anterior segments could be completely visualized in the intraoperative set up using iOCT under general anesthesia including cornea, anterior chamber angle, iris and lens despite presence of corneal opacities. iOCT was a helpful tool to detect corneal, iridal and lens-anomalies in these newborn and infants, who were not accessible to normal OCT-diagnostics without general anesthesia. Moreover iOCT enabled intraoperative decision making to perform anterior segment surgery (e.g. phototherapeutic keratectomy or lamellar keratectomy) in patients with corneal opacities. Furthermore, depth of tissue ablation could precisely be monitored during anterior lamellar keratectomy. Conclusions: Intraoperative Optical Coherence Tomography (iOCT) closes the gap of imaging diagnostics observed in newborns and infants, who have to be examined under general anesthesia and are not accessible to “routine” advanced imaging techniques so far. Furthermore iOCT proved to be a helpful tool for online monitoring of anterior segment surgery in newborn and infant children. Commercial Relationships: Sebastian E. Siebelmann, None; Philipp Steven, None; Manuel Hermann, None; Thomas Dietlein, None; Claus Cursiefen, None Program Number: 4092 Poster Board Number: B0014 Presentation Time: 8:30 AM–10:15 AM Structural and functional retinal imaging with MHz Line-field parallel swept source imaging (LPSI) Daniel Fechtig1, 2, Tilman Schmoll3, Cedric Blatter1, Rainer A. Leitgeb1, 2. 1Center for Med. Phys. and Biom. Eng., Medical University Vienna, Vienna, Austria; 2Center for Med. Phys. and Biom. Eng., Christian Doppler Society, Vienna, Austria; 33Carl Zeiss Meditec, Inc., Dublin, CA. Purpose: MHz OCT allows mitigating undesired influence of motion artifacts during retinal assessment, but comes in state-of-the-art point scanning OCT at the price of increased system complexity. By changing the paradigm from scanning to parallel OCT for in vivo retinal imaging the three-dimensional (3D) acquisition time is reduced without a trade-off between speed, sensitivity and technological requirements. Methods: Line field parallel interferometric imaging (LPSI) is utilizing a commercially available swept source, a single-axis galvo-scanner and a line scan camera for recording 3D data with up to 1MHz A-scan rate. Besides line-focus illumination and parallel detection, we mitigate the necessity for high-speed sensor and laser technology by holographic full-range imaging, which allows for increasing the imaging speed by low sampling of the optical spectrum. High B-scan rates up to 1kHz further allow for implementation of lable-free optical angiography in 3D by calculating the inter B-scan speckle variance. Results: We achieve a detection sensitivity of 93.5 (96.5) dB at an equivalent A-scan rate of 1 (0.6) MHz. Fig. 1(a) was obtained by stitching 7 slightly overlapping non-averaged tomograms. The sensitivity and resolution are high enough to visualize the external limiting membrane (ELM), and to contrast the various layers of the inner retina. Despite the loss of confocality in one lateral dimension and the shorter center wavelength of 840nm, the signal intensity from the choroid is remarkably strong maintaining structural details. In Fig. 1(b) we feature a enface projection of micro-angiographic information. One single 3D stack corresponds to a lateral FOV of approx. 5-6° and is acquired in just 2.5 s. Conclusions: Our results demonstrate for the first time competitive imaging sensitivity, resolution and speed with a parallel OCT modality. LPSI is in fact currently the fastest OCT device applied to retinal imaging and operating at a central wavelength window around 800 nm with a detection sensitivity of higher than 93.5 dB. ©2015, Copyright by the Association for Research in Vision and Ophthalmology, Inc., all rights reserved. Go to iovs.org to access the version of record. For permission to reproduce any abstract, contact the ARVO Office at [email protected]. ARVO 2015 Annual Meeting Abstracts Fig. 1(a) and 1(b) Commercial Relationships: Daniel Fechtig, None; Tilman Schmoll, None; Cedric Blatter, None; Rainer A. Leitgeb, None Support: Christian Doppler Society Program Number: 4093 Poster Board Number: B0015 Presentation Time: 8:30 AM–10:15 AM Investigation of the microscopic retina with MHz AO-OCT Omer P. Kocaoglu, Tim L. Turner, Zhuolin Liu, Donald T. Miller. School of Optometry, Indiana University, Bloomington, IN. Purpose: Optical coherence tomography (OCT) has undergone considerable technological advances over the last two decades, yet image acquisition speed remains a major limiting factor for its use in retinal imaging. Increased speed enables imaging of larger fields of view at finer spatial and temporal sampling, while also reducing the effects of eye motion. Reduction of eye motion is particularly attractive with adaptive optics OCT (AO-OCT) because the cellularlevel lateral resolution afforded by AO makes the system more susceptible to eye motion. In this study we employ a research-grade AO-OCT system that acquires images 25 times faster than clinical OCT and use it to investigate the microscopic retina in the living human eye. Methods: The Indiana MHz AO-OCT system is based on spectral domain technology. It employs a high-speed 1×4 optical switch in its novel, quad-spectrometer detection channel to achieve 1 million A-lines/s acquisition speed. The design also makes efficient use of the light available for detection. A superluminescent diode (λc=790nm, Δλ=42nm) illuminates the retina providing 5.3 μm axial resolution. The sample channel contains the AO system that dynamically corrects for ocular aberrations across a 6.7 mm pupil to provide diffraction-limited lateral resolution (1.7 μm confocal) and improved signal-to-noise ratio of retinal images. Volumes videos were acquired with the MHz AO-OCT system of all major retinal layers from the retinal pigment epithelium layer to retinal nerve fiber layer at 6 superior to the fovea. Volumes were 1.1 ×1.3 or 0.5 ×0.7 in size (containing 320 B-scans × 400 A-lines) and acquired at real time rates of 7.8 Hz and in 30 volume sequences (3.84 s). Results: MHz AO-OCT volumes were successfully acquired on three normal subjects. Retinal images were typically of ~30 dB dynamic range, without averaging, and revealed substantially reduced image blur and distortion due to eye motion compared to earlier generation AO-OCT systems developed at Indiana. The 30 dB dynamic range is comparable to that of clinical OCT and permitted visualization of all major retinal layers. The 7.8 Hz volume rate also captured the flow dynamics of retinal capillaries at different depths in the inner retina. Conclusions: MHz AO-OCT provides sufficient 3D resolution and sensitivity to image at the microscopic level any major layer of the retina. Representative MHz AO-OCT volumetric image displayed on a linear-scale. Scale bars: 25μm. Commercial Relationships: Omer P. Kocaoglu, None; Tim L. Turner, None; Zhuolin Liu, None; Donald T. Miller, #7,364,296 (P) Support: NEI grants R01-EY018339 and P30- EY019008 Program Number: 4094 Poster Board Number: B0016 Presentation Time: 8:30 AM–10:15 AM Long Working Distance Swept Source Optical Coherence Tomography for Pediatric Imaging Ruobing Qian1, Oscar Carrasco-Zevallos1, Lejla Vajzovic2, Cynthia A. Toth2, 1, Joseph A. Izatt1, 2. 1Biomedical Engineering, Duke University, Durham, NC; 2Ophthalmology, Duke University Medical Center, Durham, NC. Purpose: Optical coherence tomography (OCT) has become the standard of care for diagnosis of many retinal pathologies. However, current commercial OCT systems require cooperative patients to maintain fixation for several seconds in a chinrest. Handheld OCT systems have also been demonstrated for successful imaging of supine patients, as well as pre-term infants and neonates up to ~1 year old. However, no technology yet exists for OCT in young children due to their lack of attention and cooperation, as well as inherent fear of large objects close to their face. Therefore, we designed and built a novel OCT system with a very long working distance (distance of the last optical component of the system to the subject’s eye) to facilitate imaging of young children. Methods: A novel scanning configuration OCT system was designed to achieve a working distance of 36 cm (Fig. 1) to situate young children at a comfortable distance away during imaging. A 2f scanning configuration, instead of the conventional 4f scheme, was implemented to reduce the footprint and weight of the sample arm. To optimize optical performance at the retinal plane, the 2f system employed two custom-designed lenses (Zemax, Inc; Redmon, WA). A dichroic mirror after the objective enabled co-alignment of a LCD display used for fixation. The monitor displayed videos and targets to aid fixation during imaging.The swept-source OCT system employed a 1060 nm frequency-swept laser (Axsun Tech; Billerica, MA) and a Mach-Zender interferometer. The interferometric signal was detected with a dual-balanced receiver (Thorlabs, Inc.; Newton, NJ) and ©2015, Copyright by the Association for Research in Vision and Ophthalmology, Inc., all rights reserved. Go to iovs.org to access the version of record. For permission to reproduce any abstract, contact the ARVO Office at [email protected]. ARVO 2015 Annual Meeting Abstracts digitized at 800 MS/s (Alazar Tech Inc; QC, Canada). Custom GPUbased software enabled real-time volumetric imaging at 100,000 A-line/second. Results: The axial resolution across the 6 mm imaging depth range was measured to be 8.4um and the lateral resolution was measured to be 11-12 um. The peak sensitivity was 99.4 dB with a 3.5 mm -6dB falloff. To prove the feasibility of the system, consented adult subjects were imaged. A representative volumetric and an averaged B-scan image are shown in Fig 2. The subject was situated on a chinrest 36 cm away from the system. The LCD monitor facilitated fixation during imaging. Conclusions: A novel long working distance OCT system along with the fixation system were designed, built and tested on adult subjects. The future implication is to image the retina of young children. Commercial Relationships: Ruobing Qian, None; Oscar Carrasco-Zevallos, None; Lejla Vajzovic, None; Cynthia A. Toth, Alcon (P), Bioptigen (F), Duke University (P), Genetech (F); Joseph A. Izatt, Bioptigen (I), Bioptigen (P), Bioptigen (S) Support: The Hartwell Foundation Program Number: 4095 Poster Board Number: B0017 Presentation Time: 8:30 AM–10:15 AM Birefringence analysis of retinal nerve fiber bundles in the human eye Christoph K. Hitzenberger1, Mitsuro Sugita1, 2, Michael Pircher1, Bernhard Baumann1, Philipp K. Roberts3, Stephan Holzer3, Tomoyuki Makihira2, Nobuhiro Tomatsu2, Makoto Sato2, Clemens Vass3. 1Center F Med Physics & Biomed Eng, Medical University of Vienna, Vienna, Austria; 2Canon Inc., Tokyo, Japan; 3Ophthalmology and Optometry, Medical University of Vienna, Vienna, Austria. Purpose: To analyze birefringence, retardation, and thickness of the retinal nerve fiber layer (RNFL) along traces of retinal nerve fiber bundles in the human eye in vivo. Methods: A polarization sensitive (PS) OCT system with an integrated retinal tracker was used to record 3D data sets in healthy human eyes in vivo. En face maps of RNFL thickness, retardation, axis orientation, and birefringence were generated from the PSOCT data sets. Based on the axis orientation and thickness maps, traces of nerve fiber bundles were extracted. A new type of graphic data representation was developed: a 2D false color map that plots birefringence, retardation, and thickness along the nerve fiber bundle (y-direction) as a function of azimuthal angle (x-direction) around the optic nerve head (ONH). Based on these plots, a quantitative analysis of the variations of parameters along the nerve fiber bundle is made. Results: We analyzed the variations of retardation, thickness, and birefringence along superior and inferior nerve fiber bundles in 4 healthy eyes. If measured from the ONH outward, retardation and thickness showed a pronounced decrease along fiber traces near the ONH of > 8°/mm and > 50 mm/mm, respectively, while birefringence was rather constant, with a change of < 0.05°/mm/mm. Conclusions: In healthy eyes, RNFL birefringence is rather constant along the nerve fiber bundle length. Deviations from this pattern might be indicative of diseases like glaucoma. Nerve fiber analysis of healthy human retina from PS-OCT data set. Left: unfolded retardation map: retardation (color) is plotted along nerve fiber bundle length (y-axis) as a function of origin on azimuthal circle around ONH (x-axis). Right: unfolded birefringence map. Commercial Relationships: Christoph K. Hitzenberger, Canon Inc. (F), Canon Inc. (P); Mitsuro Sugita, Canon Inc. (E), Canon Inc. (P); Michael Pircher, Canon Inc. (F), Canon Inc. (P); Bernhard Baumann, None; Philipp K. Roberts, Canon Inc. (F); Stephan Holzer, None; Tomoyuki Makihira, Canon Inc. (E); Nobuhiro Tomatsu, Canon Inc. (E), Canon Inc. (P); Makoto Sato, Canon Inc. (E), Canon Inc. (P); Clemens Vass, Canon Inc. (P) Program Number: 4096 Poster Board Number: B0018 Presentation Time: 8:30 AM–10:15 AM Wavefront sensorless adaptive optics OCT with real time axial tracking for imaging human retina Yifan Jian1, Kevin S. Wong1, Michelle Cua1, Stefano Bonora2, Robert J. Zawadzki3, 4, Marinko V. Sarunic1. 1Engineering Science, Simon Fraser University, Burnaby, BC, Canada; 2CNR-Institute of Photonics and Nanotechnology, Padova, Italy; 3Department of Cell Biology and Human Anatomy, University of California Davis, Davis, CA; 4 Department of Ophthalmology & Vision Science, University of California Davis, Sacramento, CA. Purpose: We describe our wavefront sensorless adaptive optics optical coherence tomography (WSAO-OCT) system for imaging the human photoreceptor mosaic in vivo. Real time segmentation of the retinal OCT B-scan permitted axial tracking and extraction of en face images. We validated our system performance by imaging the retina at several eccentricities, and demonstrated the improvement in photoreceptor visibility with WSAO aberration correction. ©2015, Copyright by the Association for Research in Vision and Ophthalmology, Inc., all rights reserved. Go to iovs.org to access the version of record. For permission to reproduce any abstract, contact the ARVO Office at [email protected]. ARVO 2015 Annual Meeting Abstracts Methods: A human WSAO-OCT system was constructed using lenses (instead of mirrors), delivering a 5.5mm (diameter) at the subject’s pupil. The OCT engine acquired A-scans at a rate of 200 kHz, and volumes of 1024x200x80 voxels were acquired in ~0.1 s. A custom written GPU program tracked the layer of interest initially selected by the operator using a simple retinal segmentation algorithm, and then extracted the corresponding en face projection image. The modal WSAO algorithm optimized Zernike radial orders 2 to 4 in order to maintain a balance between optimization time and effective aberration correction. For each Zernike mode, the optimization was performed by acquiring an OCT volume and extracting an en face image for each of 5 different coefficient values. The coefficient that produced the brightest en face image was selected as the optimized value. The entire optimization process required 6~12 seconds, depending on the amount of the aberrations in the subject’s eye. Results: We present en face images acquired at three different retinal eccentricities before and after WSAO optimization in Figure 1. In the unoptimized images, the cones are mostly indistinguishable from speckle pattern. After optimization, the image contrast increased, and the cone mosaic can be resolved. Conclusions: We demonstrated a lens-based approach for WSAOOCT that is capable of resolving the cone mosaic in the human eye at small angles of eccentricity with non-mydriatic pupils even with a small-stroke DM. The reduced complexity of the lens-based WSAO design can facilitate a robust and compact imaging system that is highly suitable for clinical applications in ophthalmology. As such, correct interpretation of clinical OCT images is of great importance. Interpretation of these images, though, is clouded by optical properties of the eye, the complexity of light-tissue interactions, and image post-processing. We have designed and built a custom OCT system equipped with adaptive optics that provides greatly improved 3D resolution and permits precise instrument calibration and characterization. We investigated the OCT outer retinal band 3, thought to originate from either the cone outer segment tips or zone of interdigitation between cone outer segments (OS) and apical processes of the RPE. Methods: Five subjects were imaged between 1.5 and 4.25°. Cones were segmented and aligned by their band 3 peaks. The resulting aligned B-scan was averaged and the thickness of band 3 was measured. Next, after bulk motion correction, the phase of the band 3 reflection was extracted and variance of this phase, which is a measure of the surface’s roughness, was computed. Results: After subtracting measured axial blur, average thickness of band 3, measured in single cones, was 2.1 μm. Typical variance of bulk-motion-corrected phase measurements was 0.09 rad, which corresponds to a surface roughness of 4 nm RMS. Conclusions: Cellular measurements of band 3 suggest strongly that the reflection comes from a thin, optically smooth origin, such as a reflective surface, and not from an axially extended zone of scattering material. This surface must be located distal to the cone OS lumen and proximal to the RPE body. The most likely origin is the distal plasma membrane of the cone outer segment. Figure 1: En face images of the human photoreceptor layer acquired at several angles of eccentricity from the fovea. Commercial Relationships: Yifan Jian, (); Kevin S. Wong, None; Michelle Cua, None; Stefano Bonora, None; Robert J. Zawadzki, None; Marinko V. Sarunic, None Support: CIHR, NSERC, Michael Smith Foundation for Health Research, and Simon Fraser University Vice President of Research. NEI (R01 EY024239), UC Davis RISE Grant, NSF I/UCRC CBSS Grant. NVIDIA hardware donation program. Program Number: 4097 Poster Board Number: B0019 Presentation Time: 8:30 AM–10:15 AM Cellular morphometry of the outer retina using adaptive optics OCT Ravi S. Jonnal, Justin Migacz, Iwona Gorczynska, Robert J. Zawadzki, John S. Werner. Ophthalmology, UC Davis, Sacramento, CA. Purpose: Diseases and disorders which impact the outer retina, such as AMD, diabetic retinopathy, macular hole, and retinal detachment are routinely evaluated using optical coherence tomography (OCT). ©2015, Copyright by the Association for Research in Vision and Ophthalmology, Inc., all rights reserved. Go to iovs.org to access the version of record. For permission to reproduce any abstract, contact the ARVO Office at [email protected]. ARVO 2015 Annual Meeting Abstracts AO-OCT B-scan taken at 1.5° temporal to the fovea. In (a), band 3 reflections from single cones are marked. Axial shifts of reflections are evident. In (b), the cones are shown without axial alignment. In (c), the cones are aligned by band 3. (a) Bulk-corrected phase of band 3 of the cones in Fig. 1. Average longitudinal reflectance profiles of the (b) unaligned cones and (c) the band 3-aligned cones. Unaligned band 3 FWHM is 10.8 μm and aligned FWHM is 4.1 μm, which includes the 2.8 μm (measured) axial PSF. This suggests that the bulk of band 3’s thickness in commercial OCT images is due to axial displacements of the reflections. Commercial Relationships: Ravi S. Jonnal, US Patent No. 7,364,296; unlicensed; (P); Justin Migacz, None; Iwona Gorczynska, None; Robert J. Zawadzki, None; John S. Werner, None Support: NEI Grant R01 EY024239; unrestricted grant from Research to Prevent Blindness, Incorporated ©2015, Copyright by the Association for Research in Vision and Ophthalmology, Inc., all rights reserved. Go to iovs.org to access the version of record. For permission to reproduce any abstract, contact the ARVO Office at [email protected]. ARVO 2015 Annual Meeting Abstracts Program Number: 4098 Poster Board Number: B0020 Presentation Time: 8:30 AM–10:15 AM Visualization of Multiple Retinal Capillary Beds using Offset Pinhole Adaptive Optics Scanning Light Ophthalmoscopy Richard B. Rosen1, 2, Nadim Choudury1, 2, Nikhil Menon1, 2, Alexander Pinhas1, 2, Rishard Weitz1, Joseph Carroll3, Alfredo Dubra3, Toco Chui1, 2. 1New York Eye and Ear Infirmary, New York, NY; 2 Ophthalmology, Ican School of Medicine at Mount Sinai, New York, NY; 3Ophthalmology, Medical College of Wisconsin, Milwaukee, WI. Purpose: To image retinal capillary beds at different retinal layers in healthy and diseased retinas using an offset pinhole adaptive optics scanning light ophthalmoscope (AOSLO). Methods: Retinal capillary plexuses at different depths were imaged at various retinal locations in 2 healthy controls and 3 patients (PDR, HRVO, BRVO) using an offset pinhole AOSLO with an imaging wavelength centered at 790nm. Imaging locations on the healthy controls were located at 5° temporal, nasal, and superior to the fovea. Region of interests with vasculopathic structures were pre-identified on fundus pictures on the patients. Image sequences were acquired using 1° or 1.5° field of view at a frame rate of 15Hz. After sinusoidal distortion and eye motion were removed, averaged images and motion contrast perfusion maps were generated at different retinal depths. Results: Multiple retinal capillary beds were clearly visualized in both controls and patients using offset pinhole AOSLO. Three distinct capillary beds were resolved at all retinal locations imaged in the controls. From the 3 patients with retinopathies, vasculopathic structures such as microaneurysms and tortuous blood vessels were observed at different retinal depths. Fig 1 demonstrates a fusiform microaneurysm located in the outer retina in a patient with HRVO. This technique enabled precise depth location of normal and pathologic structures with high lateral resolution that is often a limiting factor when attempting similar stratification using optical cohernece tomography Conclusions: Offset pinhole AOSLO provides noninvasive and direct visualization of multiple retinal capillary beds at different depths, as well as the ability to isolate retinal vasculopathic structures at different retinal capillary beds. This imaging technique provides a better understanding of normal and pathological retinal vasculature development. Program Number: 4099 Poster Board Number: B0021 Presentation Time: 8:30 AM–10:15 AM Non-invasive adaptive optics imaging of the ground squirrel retina Benjamin S. Sajdak2, Christopher S. Langlo2, Yusufu N. Sulai1, Robert F. Cooper3, Dana K. Merriman5, Joseph Carroll2, 1, Alfredo Dubra1, 4 1 . Opthalmology, Medical College of Wisconsin- Eye Institute, Milwaukee, WI; 2Cell Biology, Neurobiology, & Anatomy, Medical College of Wisconsin, Milwaukee, WI; 3Biomedical Engineering, Marquette University, Milwaukee, WI; 4Biophysics, Medical College of Wisconsin, Milwaukee, WI; 5Biology, University of Wisconsin Oshkosh, Oshkosh, WI. Purpose: Ground squirrels are an important model for studying visual processing, retinal circuitry, and cone photoreceptor function. Using a custom adaptive optics scanning light ophthalmoscope (AOSLO), we sought to noninvasively image the microscopic structure of the ground squirrel retina. Methods: Four 13-lined ground squirrels (Ictidomys tridecemlineatus) were anesthetized with isoflurane (5% induction, 2-3% maintenance) and placed on a heated stage. Pupils were dilated with one drop of 2.5% phenylephrine and one drop of 1% tropicamide. Saline drops were applied as needed to maintain corneal hydration. Reflectance, confocal and split-detection AOSLO imaging through a 4 mm diameter pupil was performed with near infrared light (790 nm). The pixel size in microns was calculated using a Ronchi ruling placed in the back focal plane of a 19 mm focal length model eye, and then scaled according to the axial length of each animal, measured with an ultrasound A-scan (OTI-Scan 1000). Photoreceptor density and Voronoi geometry were calculated at known retinal locations using cell coordinates obtained by a semiautomated cell counting algorithm. Results: High-resolution images of the photoreceptor mosaic, nerve fiber layer, and retinal vasculature (Fig.1) were obtained with a high success rate (100%, n = 4). No lens opacities were observed during any of the imaging sessions, which lasted up to 1 hour. Photoreceptor density values are consistent with previously published histological data of a congener species (Long and Fisher, J Comp Neurol., 1983; Kryger et al., Vis Neurosci., 1998), ranging from 25,600–78,400 cells/mm2. The mosaic displayed a fairly triangular packing geometry, with 42-56% of the cells having six-sided Voronoi domains around the horizontal streak, and 59-69% in more peripheral locations. Conclusions: The photoreceptor mosaic, retinal capillaries and microscopic detail on the nerve fiber layer inner surface of the ground squirrel were visualized with AOSLO. The image quality and success rate appears to be better than current AOSLO efforts on mice and rats. Thus, when combined with the non-invasive high resolution imaging afforded through AOSLO, the ground squirrel could serve as a useful model to aid drug discovery and testing through longitudinal imaging on the cellular scale. Fig 1 demonstrates a fusiform microaneurysm located in the outer retina capillary layer in a patient with HRVO. The thru focus technique is able to distinguish the precise level of the lesion Commercial Relationships: Richard B. Rosen, Advanced Cellulat Technologies (C), Allergan (C), Carl Zeiss Meditech (C), Clarity (C), OD-OS (C), Opticology (I), Optovue (C); Nadim Choudury, None; Nikhil Menon, None; Alexander Pinhas, None; Rishard Weitz, None; Joseph Carroll, ImagineEyes (C); Alfredo Dubra, University of Rochester (P); Toco Chui, None Support: Marrus Foundation, Wise Foundation ©2015, Copyright by the Association for Research in Vision and Ophthalmology, Inc., all rights reserved. Go to iovs.org to access the version of record. For permission to reproduce any abstract, contact the ARVO Office at [email protected]. ARVO 2015 Annual Meeting Abstracts Ground squirrel cone photoreceptors using confocal (A) and split detector (B) AOSLO. Nerve fiber layer (C) and retinal capilaries (D). Scale bars 50mm Commercial Relationships: Benjamin S. Sajdak, None; Christopher S. Langlo, None; Yusufu N. Sulai, None; Robert F. Cooper, None; Dana K. Merriman, None; Joseph Carroll, None; Alfredo Dubra, None Support: P30 EY001931, T32-GM080202 Program Number: 4100 Poster Board Number: B0022 Presentation Time: 8:30 AM–10:15 AM Spectral domain optical coherence tomography analysis of the retina in a rat model: a comparison of transgenic immunodeficient retinal degenerate rat (SD-Foxn1 Tg(S334ter)3Lav) and rats with normal retina Alexander De Guzman1, 2, Bibo Khatib2, Robert Lin1, 2, Bryce T. McLelland2, Anuradha Mathur2, Robert B. Aramant2, Brian Cummings1, 2, Magdalene J. Seiler1, 2. 1Physical Medicine & Rehabilitation, University of California, Irvine, Irvine, CA; 2Stem Cell Research Center, University of California, Irvine, Irvine, CA. Purpose: Retinal degeneration (RD) affects millions of people worldwide, and in vivo monitoring of the progression of this disorder is needed to aid the development of treatments for it, such as retinal sheet transplants. The goal of this study is twofold: to show that retinal degeneration worsens with age, and to demonstrate that Spectral Domain Optical Coherence Tomography (SD-OCT) can track the progression of this condition in a new immunodeficient RD rat model. Methods: Retinal cross-section SD-OCT scans (Bioptigen Envisu R2200 Spectral Domain Ophthalmic Imaging System) were obtained from male and female rats (n=40) aged between P17-P77, using a group of transgenic SD-Foxn1 Tg(S334ter)3Lav rats (=RD rats) as a model for retinal degeneration, and a normal retinal control group of rats expressing human placental alkaline phosphatase (hPAP). Data analysis was done using the Bioptigen post-scan analysis software InVivoVue Diver Release 2.0 (Bioptigen, Research Triangle Park, NC). Changes in the thickness of the total retina (TR), the outer retina (OR), and the inner nuclear layer (INL) were determined with respect to age. Using SigmaPlot 11.0 (Systat Software, Inc., San Jose, CA; Fisher-corrected multiple comparison tests), layer thicknesses were compared with age group and with rat strain. Results: P17 was the earliest age in which retinas could be imaged. At this age, there was already a noticeable difference in total retinal (TR) and outer retinal (OR) thickness between hPAP rats and RD rats. In normal rats, TR was decreasing with eye growth between the ages of P17 and P77 by 10.4% (from 260μm to 233μm) whereas TR of RD rats decreased by 32.6% (from 133μm to 95μm). OR was decreasing with eye growth between the ages of P17-P77 by 6.5% (from 141μm to 132μm) whereas OR of RD rats decreased by 28.8% (from 26μm to 20μm). The INL was decreasing with eye growth between the ages of P17-P77 by 32.6% (from 38μm to 25μm) whereas the INL of RD rats decreased by 38.8% (from 34μm to 21μm). Conclusions: This study has demonstrated that SD-OCT is a viable method of detecting and analyzing retinal degeneration; and consequently, the status of the retina and any occurring changes can be monitored and analyzed over time in vivo. SD-OCT may be useful in the evaluation of treatments for retinal degeneration. Commercial Relationships: Alexander De Guzman, None; Bibo Khatib, None; Robert Lin, None; Bryce T. McLelland, None; Anuradha Mathur, None; Robert B. Aramant, Ocular Transplantation LLC (E), Ocular Transplantation LLC (P); Brian Cummings, None; Magdalene J. Seiler, Ocular Transplantation LLC (C), Ocular Transplantation LLC (P) Support: CIRM Grant TR4-06648; Summer Undergraduate Student Research Program, UC Irvine Program Number: 4101 Poster Board Number: B0023 Presentation Time: 8:30 AM–10:15 AM Google Cardboard anterior and posterior segment imaging: a valuable tool for limited-resource settings Simon Ghofrani, Mahsa Rezaei, Aaron Wang, Allen O. Eghrari, Christopher J. Brady. Wilmer Eye Institute, Johns Hopkins, Baltimore, MD. Purpose: In limited-resource settings such as developing countries, significant obstacles to ocular imaging include cost and local availability of required instruments. Smartphone photography leverages the ubiquity of mobile handsets to image the posterior pole, but typically requires the use of both hands to hold the device and a lens, which generally must be purchased separately at significant expense. Here, we describe the use of modified, open-source Google Cardboard technology to image the anterior and posterior segment using a head-mounted device composed of materials available globally and at minimal cost. Methods: A single Google Cardboard device, composed of cardboard and acrylic lenses, was modified with a strap to mount to the viewer’s head. Of the two lenses, each of which has a focal point of 4.5cm (22.2D) the right one was removed from the device and the examiner’s smartphone placed within the device to be centered in front of the left eye. A white LED light and coin cell battery were affixed to the Cardboard to maximize smartphone battery life, and installed superior to the mobile phone camera in order to provide maneuverability comparable to an indirect ophthalmoscope. Results: This device, plans for which are open source and freely available online, allows the viewer to conduct indirect ophthalmoscopy through a smartphone embedded in the headset, while recording the examination from the perspective of the examiner. The examination is facilitated with a 20D lens, but can also be demonstrated using the detached right 22.2D acrylic lens that is associated with the product, thereby allowing the examiner to ©2015, Copyright by the Association for Research in Vision and Ophthalmology, Inc., all rights reserved. Go to iovs.org to access the version of record. For permission to reproduce any abstract, contact the ARVO Office at [email protected]. ARVO 2015 Annual Meeting Abstracts perform indirect ophthalmoscopy without costly equipment. Video and clinical photographs of the technique are demonstrated. Conclusions: Using only a smartphone, cardboard and acrylic, products available in most regions of the world, indirect ophthalmoscopy can be conducted through a headset and images recorded for clinical or research use. capture human fundus images. GoPro software was then used for image processing. Results: The GoPro camera was able to capture images with both the simple eye model and the healthy human eye. Images from the study included healthy optic nerve, retinal vessel, and macular anatomy (Figure). Conclusions: The GoPro camera is capable of capturing fundus images of the human eye; however, the clinical utility of the images still needs to be demonstrated. The quality of the images rivals that of smartphone ophthalmic photography and head-to-head testing could be an area of future exploration. Additionally, while there was success with the GoPro/LED combination for coaxial camera and light source, improvements in the construct could potentially yield higher image quality. High-quality ophthalmic images obtained with relatively inexpensive and portable devices, such as the GoPro camera, will likely continue to have a growing impact on the field of ophthalmology. Still photo of indirect opthalmoscopy from video acquired through Google Cardboard. Video to be demonstrated at time of presentation reveals ease of use. Commercial Relationships: Simon Ghofrani, None; Mahsa Rezaei, None; Aaron Wang, None; Allen O. Eghrari, None; Christopher J. Brady, None Figure 1. Fundus photo of the right eye using GoPro camera device. Commercial Relationships: R Joel Welch, None; Quan Dong Nguyen, None Program Number: 4102 Poster Board Number: B0024 Presentation Time: 8:30 AM–10:15 AM A novel approach to ophthalmic photography using a portable and versatile camera device R Joel Welch, Quan Dong Nguyen. Ophthalmology, UNMC, Omaha, NE. Purpose: Smartphone fundus photography has grown in popularity over recent years. The ability to use a portable device, such as a smartphone, to capture high-resolution and clinically relevant fundus photographs has had a significant impact on patient care and interphysician communication. While the cameras of modern smartphones are growing increasingly robust, limitations do exist. To date, the GoPro camera (GoPro Inc., San Mateo, CA, USA), considered as one of the most powerful and versatile portable camera devices, has not been used in ophthalmic photography. The purpose of this study is to determine whether the GoPro camera can be used for ophthalmic fundus photography. Methods: A GoPro HERO4 Silver camera (GoPro Inc., San Mateo, CA, USA ) and a 20D lens (Volk Optical Inc., Mentor, OH, USA) were used for this study. Additionally, a battery-powered five millimeter white LED (NTE Electronics, Inc., Bloomfield, NJ, USA) was used as the coxial light source and attached to the camera using Transpore medical tape (3 M, St. Paul, MN, USA). A simple and inexpensive model of the human eye was constructed to aide the author in learning image capture technique before approaching a human volunteer. Once the technique was mastered using the model, the right eye of one human volunteer was dilated in order to safely Program Number: 4103 Poster Board Number: B0025 Presentation Time: 8:30 AM–10:15 AM Slitlamp-mounted smartphone adapters using reverse engineering with 3D scanning and 3D printing Charline Boente1, Ethan Tu2, Rony Sayegh1. 1Ophthalmology, University Hospitals Case Medical Center, Cleveland, OH; 2 Biomedical Engineering, Case Western Reserve University, Cleveland, OH. Purpose: Slitlamp smartphone adapters have become widely available, however, only a limited number of phone models are supported. We describe a process using 3D scanning and 3D printing to build customizable adapters that fit virtually any smartphone to any slitlamp. Methods: With the selected smartphone, we obtained precise dimensions of the phone using 3D scanning (NextEngine 3D Scanner, Santa Monica, CA), which outputs a stereolithography (STL) file that can be manipulated in a variety of design software programs. We used AutoCAD software (AutoDesk, Inc. San Rafael, CA) to design a custom adapter from the dimensions obtained from the scanned phone using reverse engineering principles. A prototype of the custom adapter was then produced using 3D printing (Stratasys Fortus 400mc, Eden Prairie, MN). Each customized model was tested on our clinics’ slitlamps and verified for fit and ease of obtaining photographic images of the anterior segment. Results: Slitlamp-mounted adapters for a variety of iPhone (Apple, Cupertino, CA) models as well as the Samsung Galaxy S4 (Samsung Electronics Co. Ltd., Ridgefield Park, NJ) were built using this ©2015, Copyright by the Association for Research in Vision and Ophthalmology, Inc., all rights reserved. Go to iovs.org to access the version of record. For permission to reproduce any abstract, contact the ARVO Office at [email protected]. ARVO 2015 Annual Meeting Abstracts method. Precise measurements were obtainable using 3D scanning which were used to build accurate and custom adapters using AutoCAD followed by 3D printing. Conclusions: 3D scanning provides easy data acquisition of the phone dimensions that improves the accuracy of the 3D printed adapters and allows a wider range of phone models to be used for slitlamp photography. Commercial Relationships: Charline Boente, None; Ethan Tu, None; Rony Sayegh, None Program Number: 4104 Poster Board Number: B0026 Presentation Time: 8:30 AM–10:15 AM Smartphone fundus photography, in vivo retinal fluorescent photography and fluorescein angiography in mouse eyes Cynthia X. Qian1, Eiichi Hasegawa1, Luis Haddock2, David M. Wu1, 3, Shizuo Mukai1. 1Ophthalmology, Massachusetts Eye and Ear Infirmary, Boston, MA; 2Bascom Palmer Eye Institute, Palm Beach Gardens, FL; 3Genetics, Harvard Medical School, Boston, MA. Purpose: To describe a technique of fundus photography, in vivo fluorescent retinal photography and fluorescein angiography in mouse eyes using a smartphone Methods: Wild-type mice with normal fundi and with retinal detachment and retinas expressing Green Fluorescent Protein (GFP) or Discosoma Red Fluorescent Protein (DsRed) secondary to adeno-associated virus (AAV) infection or electroporation. After pharmacologic mydriasis, the mice were gently held in hand without anesthesia. Using a modification of our techniques previously described for human and rabbit eyes (Haddock et al. J Ophthalmol 2013, Article ID 518479) fundus photography was carried out using an iPhone 5s® or iPhone 6® and a 78D lens (as compared to 20D and 28D lenses for human and rabbit eyes respectively). For in vivo fluorescent photography of retinas transduced with GFP or DsRed, an excitation filter (but no barrier) for fluorescein angiography from an old fundus camera was placed in front of the LED light source but not the camera lens of the iPhone®. For fluorescein angiography, an excitation filter was place in front of the LED and a barrier filter was placed in front of the camera lens, and images were taken after intraperitoneal injection of fluorescein. Results: Using this system we were able to consistently take high-quality video clips and fundus photographs in the eyes of awake mice. The use of an excitation filter allowed for fluorescent photography of the patches of subretinal GFP and DsRed transgene expression. Using a combination of excitation and barrier filters we were able to perform fluorescein angiography in wild-type mice. Conclusions: We demonstrated that with a modification of a previously described system for human and rabbit eyes, fundus photography, in vivo retinal fluorescent photography and fluorescein angiography can be performed using the iPhone®. This technique is relatively inexpensive, readily available, and very portable. Fluorescein angiography imaging highlighting normal vasculature in a mouse retina Fluorescent photography highlighting patch of DsRed staining in a rd1 mouse retina Commercial Relationships: Cynthia X. Qian, None; Eiichi Hasegawa, None; Luis Haddock, None; David M. Wu, None; Shizuo Mukai, None ©2015, Copyright by the Association for Research in Vision and Ophthalmology, Inc., all rights reserved. Go to iovs.org to access the version of record. For permission to reproduce any abstract, contact the ARVO Office at [email protected]. ARVO 2015 Annual Meeting Abstracts Program Number: 4105 Poster Board Number: B0027 Presentation Time: 8:30 AM–10:15 AM Comparison of Smartphone Ophthalmoscopy with Slit-lamp Biomicroscopy for Undilated Glaucoma Screening Andrea Russo1, Francesco Morescalchi1, Luisa Delcassi1, Ciro Costagliola2, Francesco Semeraro1. 1Univ degli Studi di Brescia Italy, Brescia, Italy; 2Eye Clinic, University of Molise, Campobasso, Italy. Purpose: Smartphone ophthalmoscopy is a recent and promising technique to screen the community. The purpose of this study is to evaluate the agreement of smartphone ophthalmoscope and slitlamp biomicroscopy to discriminate eyes with glaucoma from those without. Methods: One hundred glaucoma patients and 100 healthy control subjects were included. All of them underwent visual field test and undilated smartphone ophthalmoscopy with D-Eye device (Si14 S.p.A., Padova, Italy) followed by dilated optic nerve head (ONH) slit-lamp examination. The following parameters were evaluated: cup/disc ratio; neuroretinal rim pallor (4-step scale); peripapillary chorioretinal atrophy; and optic disc hemorrhages. Results: All included eyes completed the study. Cup/disc ratio exact agreement between the two methods was observed in 186 (93%) of 200 eyes (simple κ = 0.91; 95% CI 0.87–0.95) and agreement within one step was observed in 200 eyes (100%). Rim pallor exact agreement was observed in 179 eyes (89.5%; simple κ = 0.85; 95% CI 0.79–0.91) and agreement within one step was observed in 197 eyes (98.5%). An almost perfect agreement was observed for both peripapillary chorioretinal atrophy and optic disc hemorrhages. Conclusions: Undilated smartphone ophthalmoscopy showed a very good agreement with dilated biomicroscopy for ONH assessment. Smartphone ophthalmoscopy can accurately detect ONH parameters and might be used as a screening tool for glaucoma. Representative retinal images of optic nerve head taken with D-Eye. (A) Cup/disc ratio evaluated as 0.6 in a 76-year-old male patient with nuclear cataract. (B) Healthy optic nerve head in a 23-year-old female. (C) Depiction of the D-Eye prototype magnetically attached to the smartphone. Commercial Relationships: Andrea Russo, None; Francesco Morescalchi, None; Luisa Delcassi, None; Ciro Costagliola, None; Francesco Semeraro, None Program Number: 4106 Poster Board Number: B0028 Presentation Time: 8:30 AM–10:15 AM Smart phone retinal photography: Uses and experiences with the novel retinal imaging technique Luis Leon, Jonathan I. Huz, Arkadiy Yadgarov, Marco A. Zarbin, Neelakshi Bhagat. Ophthalmology, Rutgers New Jersey Medical School, South Orange, NJ. Purpose: Retinal imaging is an essential component in the practice of ophthalmology; its use is limited to the availability of necessary imaging equipment. We reviewed fundus photos taken using a recently described technique of fundus photography using a smartphone, and its utility in a large hospital with a busy ophthalmology training program. This study describes and evaluates the use of this technique in a setting when fundus cameras are not available. Methods: We performed a retrospective review of all fundus photographs of retinal pathology taken using smart phone cameras in the inpatient or emergency room settings. Fundus images were captured with an iPhone 4, 5 or 6 and a 20D lens. Using the coaxial light source from the phone and its included video software,the phone functions as an indirect ophthalmoscope, high-definition videos of the fundus can be recorded for subsequent still image extraction. The pictures were evaluated by ophthalmologists to asses its quality and clinical utility. Results: We obtained fundus photos of patients with retinal pathology incluiding retinal detachment, pre-retinal membranes, proliferative diabetic retinopathy with pre-retinal and vitreous hemorrhage, macular pucker, commotio and retinal hemorrhages of shaken baby syndrome . Retina specialists the residents easily identified the pathology that was photographed using this technique. Pictures were of very good quality even when taken by ophthalmology residents that were inexperienced with the technique. The resolution and details observed in the photos was sufficient to recognize important retinal details and to establish a diagnosis. The pictures were also noted to be an excellent resource to compare disease progression and changes of retinal anatomy in subsequent examinations. Conclusions: In our review, we noted that the technique to obtain retinal pictures with smartphones is an excellent resource, especially in the emergency room and inpatient setting. With this technique eye doctors can assess and follow up retinal pathology effectively, high quality fundus photos can be obtained by relatively inexperienced personnel. This technique should be introduced to all eye care specialists in order to effectively document retinal findings when there are no retinal cameras available; this technique could also be introduced to emergency room personnel in order to evaluate a patient’s retina with minimal equipment cost. Commercial Relationships: Luis Leon, None; Jonathan I. Huz, None; Arkadiy Yadgarov, None; Marco A. Zarbin, None; Neelakshi Bhagat, None Program Number: 4107 Poster Board Number: B0029 Presentation Time: 8:30 AM–10:15 AM The Potential Positives of Smartphone Photography Using Negative Images David L. Nash, Eric Crouch, Shannon McCole. Ophthalmology, Eastern Virginia Medical School, Norfolk, VA. Purpose: Smartphone photography has become increasingly popular and has several applications. We retrospectively reviewed and ©2015, Copyright by the Association for Research in Vision and Ophthalmology, Inc., all rights reserved. Go to iovs.org to access the version of record. For permission to reproduce any abstract, contact the ARVO Office at [email protected]. ARVO 2015 Annual Meeting Abstracts compared standard color photographs with the negative photographic image of several retinal findings photographed with a handheld smartphone to identify any utility in the use of negative retinal images with smartphone photography. Methods: Using an iPhone 4S (Apple Inc., Cupertino, CA, USA) and a handheld 20 or 28 diopter lens to capture retinal images of 4 separate patients with a variety of retinal conditions seen by a single resident at a metropolitan based, residency program, color photographs were compared to the negative images. The negative images were created with the application Negative Me (Fuen Mao © 2010-2013 1 stfancy Studio). The negative images were compared retrospectively to their color counterparts. Photographs were cropped to remove patient identifying information. All patients signed hospital consent forms to be photographed. Results: Smartphone negative retinal images, when compared to their color image, best highlight hypopigmented lesions (such as Cytomegalovirus (CMV) scarring), lightly colored structures on the retina such as the optic disc, and areas or retinal pigment epithelium loss. Examination of negative images taken three weeks apart of a patient with CMV retinitis may highlight areas of retinal scarring. Conclusions: Smartphone fundoscopy has many applications. Negative images are simple to create using smartphone technology. Certain conditions, most notably those involving hypopigmentation of the retina, may benefit from the use of negative imagery. There may also be utility in evaluating cup to disc ratios with negative imaging. This study acts as a pilot study. Further study is needed to validate the utility of negative imagery and its clinical application in disease management. Image 1: a: Color photograph taken with a 20 diopter lens. b: Color photograph taken with a 28 diopter lens. c: Negative image of 1a. d: Negative image of 1b. All images are inverted. Image 2: a: Color photograph taken with a 20 diopter lens. b: color photograph taken 3-4 weeks later of the same eye with a 20 diopter lens. c: Negative image of 3a. d: Negative image of 3b. All images are inverted. Commercial Relationships: David L. Nash, None; Eric Crouch, None; Shannon McCole, None Program Number: 4108 Poster Board Number: B0030 Presentation Time: 8:30 AM–10:15 AM Wide-Field Bedside Imaging of the Human Retina Using a Smartphone Adapter Lianna Valdes, Dov B. Sebrow, Tongalp H. Tezel. Edward S. Harkness Eye Institute, Department of Ophthalmology, Columbia University College of Physicians and Surgeons, New York, NY. Purpose: To develop and evaluate the clinical utility of a smartphone-based portable fundus camera that can be used for widefield imaging and recording of fundus pathologies at bedside. Methods: A polyurethane mount was designed and constructed to keep the camera module of a smartphone (iPhone 4S) aligned with the assistant mirror of an indirect ophthalmoscope (Keeler Inc., Broomall, PA). Pretesting was performed to determine the optimal lighting and focusing conditions for video and single-shot photography of the human fundus. Next, macular and peripheral images of various fundus pathologies were obtained using both a standard fundus camera (Zeiss FF450, Dublin, CA) and the smartphone-based camera. The clarity of the images, correct identification of the pathologies, and ease of obtaining and storing the fundus images were compared using 8 masked ophthalmologists. A 3D printable model of the camera mount was created with a 3D scanner. Results: High-quality imaging of the posterior pole and retinal periphery was achieved using the novel iPhone-adapted indirect ophthalmoscope. Best results were obtained when positioning the iPhone camera 0.5 cm from the assistant mirror and using medium intensity illumination (~0.6 mW cm-2 sr-1) from the indirect ophthalmoscope. When looking at the fundus images captured by the smartphone camera, survey participants accurately described the details of the fundus pathology 72% of the time. After viewing ©2015, Copyright by the Association for Research in Vision and Ophthalmology, Inc., all rights reserved. Go to iovs.org to access the version of record. For permission to reproduce any abstract, contact the ARVO Office at [email protected]. ARVO 2015 Annual Meeting Abstracts the corresponding standard fundus photos, only 28% of respondents changed their initial diagnosis correctly. The iPhone camera was easy to use and was able to image far peripheral lesions that were located beyond the capture range of the stationary fundus camera. It allowed documentation of a peripheral retinal lesion during dynamic depression. It also has the advantage of being considerably less expensive and more portable than other modalities. Conclusions: The Smartphone-adapted indirect ophthalmoscope can provide clinically useful imaging of the posterior pole and retinal periphery. This efficient, cost-effective, and portable imaging modality will allow rapid acquisition, storage, and transmission of fundus images in the pediatric population and in patients unable to sit for standard fundus photography. We believe it will be especially useful for documenting and monitoring the fundus changes in retinopathy of prematurity. Commercial Relationships: Lianna Valdes, None; Dov B. Sebrow, None; Tongalp H. Tezel, None Support: Columbia University College of Physicians and Surgeons Scholarly Projects Program Program Number: 4109 Poster Board Number: B0031 Presentation Time: 8:30 AM–10:15 AM AMD - A Metamorphopsia Detector Daniela Claessens1, Ronald V. Krüger2. 1Augenärztliche Gemeinschaftspraxis Lindenthal, Cologne, Germany; 2app4eyes, Meinerzhagen, Germany. Purpose: Purpose: We developed the computer based App AMD -A Metamorphopsia Detector- to provide a self-control monitoring tool to detect and control the degree and size of metamorphopsias and scotomas. This study was performed to examine sensitivity and specificity of AMD and its correlation with anatomical course, functional development and quality of life. Methods: Methods: AMD is based on the Amsler grid. The software uses the concept of a negative image: a distorted image can be straightened by moving the mouse. Degree and dimension of distorted lines or scotoma are transformed into indices. I) In a pretest sensitivity and specificity of the AMD App were examined in 17 healthy and 5 eyes with metamorphopsias due to macular degeneration (age 61-83). II) In a second step 19 eyes (age 24-91) with maculopathies performed the metamorphopsia module alone or in combination with the scotoma module. III) In a prospective observational clinical pilot study we examined monocular best corrected visual acuity (BCVA), Amsler grid, vision related quality of life (National Eye Institute Visual Function Questionnaire NEI VFQ 25), OCT (optical coherence tomography) and AMD in 13 eyes (age 37-91) with macular edema before and after intravitreal injections of anti vascular endothelial growth factor. Prior to the study all patients signed informed consent. Results: Results: I) Pretests confirmed construct validity and reliability. Sensitivity of AMD was 80%, specificity was 100%. Chi2-Test lead to rejection of the zero hypothesis (c2 > 3.84; freedom degree = 1, α = 0.05). II) Including a wider range of pathologies, addition of the scotoma module increased sensitivity from 0.84 to 0.94. OR to detect macular pathologies without metamorphopsias was 3.37; 95% CI: 0.94 - 5.67 when combining both app modules. III) a-VEGF improved OCT and AMD in all 13 eyes: average decrease of macular thickness was 77.54 mm (median 55 mm; SD 49.99; CI 104.75; 50.33) and 6.0 (median 4.76; SD 3.5; CI 7.9; 4.1) for metamorphopsia index. Mean gain in NEI VFQ 25 was 8.77 (median 19; SD 12.39; CI 15.51; 2.03).BCVA improved in 10 and was stable in 3 patients. Geometric mean BCVA was 0.38 logMAR (SD 0.28) before and 0.25 logMAR (SD 0,21) after treatment. Conclusions: Conclusions: As a quantitative test AMD can support conventional diagnosis, lower the threshold for diagnosis,enhance compliance and adherence in the treatment of age related macular degeneration and other diseases leading to macular edema. Commercial Relationships: Daniela Claessens, None; Ronald V. Krüger, app4eyes (I) Program Number: 4110 Poster Board Number: B0032 Presentation Time: 8:30 AM–10:15 AM Smart Ophthalmics: A Smart Service Platform for TeleOphthalmology Wolfgang Fink, Mark Tarbell. Vis & Autonomous Explorat’n Sys, University of Arizona, Tucson, AZ. Purpose: To provide ophthalmic healthcare to people who are geographically dispersed (e.g., rural populations), or operate/ live in austere environments (e.g., military, third world, natural disaster), where time, cost, and possibility of travel make access to even adequate medical care difficult. As a result, significant causes of preventable vision loss, such as ocular trauma, glaucoma, and macular degeneration, may be detected early and treated in time to prevent permanent vision impairment. Methods: Optical extension devices are attached to a smartphone to turn it into a mobile ophthalmic examination device. A custom app is engaged to perform an eye exam specific to the attached optical extension. Using a wireless connection, the smartphone app submits the collected examination data to a remote “expert system,” which provides medical analysis. The analysis results are sent back to and displayed on the smartphone. Results: A server-based telediagnostic analysis capability for current and future smartphone-based ophthalmic examination devices has been developed. Examination data, gathered with such devices, can now be sent via WiFi or cell signal through a smartphone app to a server for automated analysis, the results of which are sent back to the originating smartphone almost immediately. As a first analysis example, the cup-to-disc ratio in fundus images is determined to assist the early detection of glaucoma. The fundus images are ©2015, Copyright by the Association for Research in Vision and Ophthalmology, Inc., all rights reserved. Go to iovs.org to access the version of record. For permission to reproduce any abstract, contact the ARVO Office at [email protected]. ARVO 2015 Annual Meeting Abstracts obtained via a commercial-off-the-shelf handheld ophthalmoscope that is attached to a smartphone. Moreover, a smartphone-based miniaturized pupillometer has been developed that allows for the recording of pupillary behavior in the presence and absence of light stimulation, i.e., pupillary light reflex and dark reaction. The pupillary diameter as a function of time is displayed in addition to parameters such as latency, constriction, and dilation times. Conclusions: Smartphone-based mobile ophthalmic examination devices combined with server-based smart telediagnosis capabilities pave the way towards smart ophthalmics, i.e., smart tele-ophthalmology. This paradigm may greatly improve remote patient screening and triage. It may help ensure that patients with undiagnosed eye diseases are detected early and treated in time to prevent permanent vision impairment. Establishing this paradigm has the potential to change the very economy of extending quality healthcare to many while reducing cost. Commercial Relationships: Wolfgang Fink, California Institute of Technology (P), University of Arizona (P); Mark Tarbell, University of Arizona (P) Support: NSF Award #IIP-1430062 Program Number: 4111 Poster Board Number: B0033 Presentation Time: 8:30 AM–10:15 AM Mobile application software (App) for Prediction of Advanced Age-related Macular Degeneration Min-Lee Chang1, Yu-Hsuan Chiu3, Chung-Jung Chiu1, 2, Luca Avoni4. 1USDA Human Nutrition Research Center on Aging, Tufts University, Boston, MA; 2Department of Ophthalmology School of Medicine, Tufts University, Boston, MA; 3Concord-Carlisle High School, Concord, MA; 4Department of Ophthalmology, Bologna Hospital Ospedale Maggiore, Bologna, Italy. Purpose: To develop a state-of-the-art mobile application software (App) for the macular risk scoring system (MRSS). Methods: The MRSS algorithm uses information from a patient’s age, sex, race, education level, smoking status, macular pigmentation (normal vs. abnormal), maximal drusen size, and drusen texture (hard vs. soft) to calculate a risk score for progression to advance agerelated macular degeneration (AMD). Based on a one-page prototype of the MRSS App for the Age-Related Eye Disease Study (AREDS) that have been developed and published along with the MRSS paper (Chiu et al., Ophthalmology 2014;121:1421-1427.), we wrote a userfriendly App for iOS devices (iPhones/iPads) and Android devices and submitted the App to the Apple App Store and the Google Play, respectively. Results: The App consists of 12 pages, including a cover page, a welcome page, a credit page, 8 question pages for the eight predictors, and a result page that automatically presents the result from the MRSS. Before submitted for download, the App was tested to ensure consistent results with both of the prototype of the App and manual calculation from the MRSS algorithm. The App is free for download at https://itunes.apple.com/us/app/macular-degenerationtest/id944048670?l=it&ls=1&mt=8 for iOS devices and at https:// play.google.com/store/apps/details?id=com.arscolor.app.mrss&hl=it for Android devices. Conclusions: We have refined the AREDS MRSS App prototype and made the App available for download via Internet. To our knowledge, our App is the first mobile application for the prediction of advanced AMD. Based on the results from the MRSS paper, this App can serve as a platform for modifications to work in other populations. Commercial Relationships: Min-Lee Chang, None; Yu-Hsuan Chiu, None; Chung-Jung Chiu, None; Luca Avoni, None Program Number: 4112 Poster Board Number: B0034 Presentation Time: 8:30 AM–10:15 AM A New iPad and iPhone App to Manage Patients with Dry Eye caterina cordella1, Edoardo Oliveri3, Massimo Sonnati3, Stefano Barabino2. 1Farmigea SpA, Pisa, Italy; 2Clinica Oculistica, DiNOGMI, Azienda Ospedaliera San Martino-IST, Genoa, Italy; 3 Hippocrates Sintech, Genoa, Italy. Purpose: Dry eye syndrome (DES) is a disease of the ocular surface and one of the most frequent pathological conditions in ophthalmology. The aim of the project was to develop an app to offer the possibility to use iPad and iPhone to record patients’ complete clinical chart data with a specific section dedicated to ocular surface tests and DES management that could be used in everyday clinical practice and for clinical studies. Methods: In the first phase we compared numerous clinical charts available for PC and paper versions. Then we developed a complete clinical chart (iEyes) to record patients’ picture, medical and ophthalmological history, visual acuity, and clinical data for iPad. In particular we have included a section with specific ocular surface tests: dry eye questionnaires, Schirmer test, fluorescein and lissamine green staining grading scales, tear break-up time, conjunctival hyperemia score, eyelid margin redness, quality of expressed meibum, and other specific tests used in experimental studies only. The app was tested by 3 general ophthalmologists and 3 specialists in ocular surface diseases. Data were recorded with standard methods (paper) and with the new app. Recording time were measured and compared. Data could visualized on i Phone also. Results: A total of 300 patients were recorded, with ocular surface tests performed in 90 patients. No problems were referred by physicians with the use of the app on iPad. The recording time was significantly (P<0.05) lower in the iPad group compared to the paper group. The chance to visualize clinical chart recorded data on iPhone obtained a significant satisfaction rate. Conclusions: This new app provides a useful tool for clinicians and researchers to record data that could be used in everyday practice and for clinical studies. Future developments could be the possibility for patients included in clinical studies to communicate data of questionnaires from their devices and to verify treatment compliance. Commercial Relationships: caterina cordella, Farmigea SpA (E); Edoardo Oliveri, Hippocrates Sintech (E); Massimo Sonnati, Hippocrates Sintech (E); Stefano Barabino, None ©2015, Copyright by the Association for Research in Vision and Ophthalmology, Inc., all rights reserved. Go to iovs.org to access the version of record. For permission to reproduce any abstract, contact the ARVO Office at [email protected]. ARVO 2015 Annual Meeting Abstracts Program Number: 4113 Poster Board Number: B0035 Presentation Time: 8:30 AM–10:15 AM Automated ocular surface staining analysis based on a teachable algorithm: smartphone web-based applicayion for dry eye and ocular surface disease Felipe H. Padovani1, Paulo Borges5, Kristopher Sanford5, Luciana Oharomari4, Eduardo M. Rocha4, Paulo Schor3, Monica Alves2. 1 Pontific Catholic University of Campinas, Jundiaí, Brazil; 2 Ophthalmolgy, University of Campinas, Campinas, Brazil; 3 Ophthalmology, Federal University of São Paulo, São Paulo, Brazil; 4Ophthalmology, FMRP, Ribeirão Preto, Brazil; 5Electrical Engineering, Santa Clara University, Santa Clara, CA. Purpose: Dry Eye Disease (DED) and ocular surface disorders diagnosis, scoring and follow up apply vital stainings. However, interpretation is influenced by observer experience and subjectivity. The aim of the present work is to describe a web-based application (app) which runs an algorithm to analyzes images of fluorescein ocular surface staining obtained with the digital camera of a smartphone to quantify, register and provide a follow up tool. Methods: Support Vector Machine (SVM) was applied to score the intensity and pattern of fluorescein staining in images of ocular surface obtained from healthy individuals and ocular surface diseases patients using the camera of the smartphone connected to a slit lamp. We are trying an automated approach to reduce the noise and to crop the unwanted background, using a sequence of algorithms from the Open Computer Vision (OpenCV) programming library. The goal is to obtain an image that can be analysed by a standard method which produces a numerical coefficient tha can be compared to clinical results. Results: The software determines the optimal hyperplane to provide the most accurate maximum margin to separate the staining area from not one, as seen in punctate keratitis in DED, corneal ulcers and epithelial defects. Gauss filters are being applied to improve the final results. Preliminary results have showed necessity to improve the techniques in order to reduce the noise and crop the image properly and, then, improve the statistical and clinical relevance of the numeric coefficient. Conclusions: The present work describes a new tool capable to register and score corneal fluorescein staining. It will be useful for clinical standardization and follow up measurements of DED severity and ocular surface disorders. Regarding practical aspect this tool can run in devices present in the daily clinical practice, the slit lamp and the smartphone, providing a feasible and more precise method to analyse ocular surface staining. Normal ocular surface: fluorescein staining under cobalt filter, smartphone and slit lamp photograph 10x magnification. Image editing algorithm prior to staining analysis. Commercial Relationships: Felipe H. Padovani, None; Paulo Borges, None; Kristopher Sanford, None; Luciana Oharomari, None; Eduardo M. Rocha, None; Paulo Schor, None; Monica Alves, None Program Number: 4114 Poster Board Number: B0036 Presentation Time: 8:30 AM–10:15 AM Contribution of augmented reality in clinical practice in ophthalmology Mathieu Flores1, Nina Mauris2, Anne-Charlotte Level2, Julien Magnin-Feysot2, Maher Saleh1. 1CHU Besançon, Besançon, France; 2 ISIFC, Besançon, France. Purpose: Feasibility study of a device allowing recording, transmission and viewing by another operator of a stereoscopic clinical examination at the slit lamp. Methods: Two medical cameras (CM900, Haag-Streit, Bern, Switzerland) were mounted on a slit lamp (BQ900, Haag-Streit, Bern, Switzerland) equipped with a beam splitter allowing to record stereoscopic videos. The video footage associated with the patient’s clinical data were then remotely transmitted to another examiner located in another room via a secure server dedicated to telemedicine. The data was then visualized through augmented reality glasses type ©2015, Copyright by the Association for Research in Vision and Ophthalmology, Inc., all rights reserved. Go to iovs.org to access the version of record. For permission to reproduce any abstract, contact the ARVO Office at [email protected]. ARVO 2015 Annual Meeting Abstracts Oculus Rift DK2 (Oculus VR, California) for stereoscopic viewing of the recorded consultation. Results: Five patients consulting for diseases of the anterior and posterior segments were examined remotely and the diagnosis was consistent with that of the first examiner in all cases. Conclusions: The success of this experience of telemedicine and augmented reality offers significant opportunities in the fields of education and remote consultation. Commercial Relationships: Mathieu Flores, None; Nina Mauris, None; Anne-Charlotte Level, None; Julien Magnin-Feysot, None; Maher Saleh, None Program Number: 4115 Poster Board Number: B0037 Presentation Time: 8:30 AM–10:15 AM A novel imaging technique to non-invasively track molecules in the eye Guillaume Normand1, Jan Penraat2, Joy Ghosh4, Kellyann Kovach2, Mikele Simkins2, Michael Rosol1, Cynthia L. Grosskreutz3, Sudeep Chandra1. 1BioMarker Development, Novartis Institutes for Biomedical Research, East Hanover, NJ; 2LAS, Novartis Institutes for Biomedical Research, East Hanover, NJ; 3Translational Medicine, Novartis Institutes for Biomedical Research, Cambridge, MA; 4 Ophthalmology Research, Novartis Institutes for Biomedical Research, Cambridge, MA. Purpose: Understanding biodistribution in the eye is a critical step in developing novel therapeutics, yet outside of small animals the ability to visualize such distribution remains limited. A novel in vivo imaging method was evaluated to non-invasively and longitudinally measure the distribution of molecules after intravitreal (IVT) injection. The goals of this study were to select the optimal dye for in vivo use, to build a quantification method and to develop a translatable technology. Methods: The near-infrared probes IRDye800CW and Indocyanine Green (ICG), as well as fluorescein, were evaluated ex vivo and in vivo after IVT or intravenous (IV) dosing. Cynomolgus monkeys were taken to the imaging room under anesthesia. After pupil dilation, animals were placed in the prone position and images were captured with a Spectralis, a confocal laser ophthalmoscope (cSLO), using 30 degree and ultra-widefield lenses (UWF) and at different sensitivities and diopters. Fluorescence intensity was measured at the retina and in the vitreous using the CellProfiler software. Results: IRDye800CW was found to be brighter both ex vivo and in vivo and less prone to quenching at high concentrations as compared to ICG. Moreover, IRDye800CW was shown to be cleared less rapidly than fluorescein and ICG from the retina and blood pool after IVT or IV injection, respectively. Based on a dose comparison study, the minimum detectable concentration of IRDye800CW at the retina was determined to be between 0.0349ug and 0.349ug. After IVT injection of 3.49ug of IRDye800CW, fluorescence intensity at the retina peaked 24h post-dose but was undetectable after 15 days post-dose. Interestingly, IRDye8000CW was observed to accumulate in the optic disc as well as form speckles along the retinal arteries. The UWF lens was able to demonstrate the high variability of the injection site and pattern by capturing a larger area of the retina. Conclusions: Of the dyes tested, IRDye800CW was validated in this study as the optimal dye for in vivo ocular imaging and potential future clinical use. Distribution of the free dye was measured in several animals in order to benchmark the passage of the free dye as compared to other targeted probes. In contrast to other methods, this is a non-invasive technique that combines qualitative as well as semi-quantitative measurements of a molecule’s vitreous passage and provides useful anatomical information, especially for the retina. Commercial Relationships: Guillaume Normand, Novartis (E); Jan Penraat, Novartis (E); Joy Ghosh, Novartis (E); Kellyann Kovach, Novartis (E); Mikele Simkins, Novartis (E); Michael Rosol, Novartis (E); Cynthia L. Grosskreutz, Novartis (E); Sudeep Chandra, Novartis (E) Program Number: 4116 Poster Board Number: B0038 Presentation Time: 8:30 AM–10:15 AM In vivo low light level mouse retinal SLO imaging: limits for backscattered intensity and autofluorescense detection Pengfei Zhang1, Azhar Zam1, Edward Pugh1, Robert J. Zawadzki1, 2. 1 Cell Biology and Human Anatomy, University of California, Davis, Davis, CA; 2Dept. of Ophthalmology & Vision Science, University of California, Davis, Sacramento, CA. Purpose: To evaluate feasibility of a custom mouse scanning light ophthalmoscope (SLO) for low light level (below 1mW) mouse retinal imaging. Low light imaging is desired to reduce potential deleterious effects of light-tissue interactions, including heating and photochemical reactions (e.g., reactive oxygen species; retinoid byproducts from pigment bleaching). Methods: A custom widefield (~50 deg VA) high-resolution retinal imaging SLO with a 0 Dpt contact lens (affixed to the SLO optics) was used to simultaneously acquire both backscattered intensity as well as autofluorescense signal from mouse retina. Different scanning speeds and light levels were tested to find the minimum light power that gives good quality images. 469 nm light from a Fianium sc400 Supercontinuum laser source was used for imaging and a 503 nm long-pass filter was used to select retinal autofluorescense. The SLO detection system consisted of a Hamamatsu PMT (H742220) for reflected light and H7422-40 for fluorescense and Femto preamplifiers (107 gain), and the sampling rate is 500kHz. Custom Labview acquisition software allowed selection of the sampling region (ROI) and frame rate. Pigmented (C57BL/6) and albino (BALB/cJ) mice from Jackson Laboratory were used as subjects. During imaging, mice were anesthetized with 2-3% inhalational isoflurane anesthetic. Pupils were dilated and cyclopleged with Tropicamide and phenylephrine. Results: Example images of mouse retina (BALB/cJ, 13 mo) acquired simultaneously in reflectance and fluorescence channels during a single imaging session with different light levels are presented (Fig. 1). The data were acquired with pixel sample time of 6 μs and sampling density of 256 x 256 pixels, resulting in frame rate of ~2Hz. Control gain voltage of the PMTs was adjusted to match the dynamic range of the DAQ during imaging. Conclusions: Low light level mouse retinal imaging is essential to allow study of physiological processes of the retina in vivo without adversely affecting retinal structure and function by the probing beam itself. Our data suggest that low light level SLO is feasible for mouse retinal imaging. In particular, light levels below 1mW can be used during mouse positioning, keeping light exposure to minimum. ©2015, Copyright by the Association for Research in Vision and Ophthalmology, Inc., all rights reserved. Go to iovs.org to access the version of record. For permission to reproduce any abstract, contact the ARVO Office at [email protected]. ARVO 2015 Annual Meeting Abstracts Conclusions: The translation of this approach into clinical practice would enable visualisation of immune cells in situ. This will not only provide a greater understanding of pathogenesis, monitoring and assessment of therapy in many human ocular diseases but also opens the ability to image immunity live for neurodegenerative disorders, cardiovascular disease and systemic immune mediated disorders. Fig.1 Single frame and 50 frames summed reflectance(REFL) and autofluorescence(AF) images with different light levels. Commercial Relationships: Pengfei Zhang, None; Azhar Zam, None; Edward Pugh, None; Robert J. Zawadzki, None Support: UC Davis Research in Science & Engineering (RISE) grant, UC Davis National Eye Institute Core facilities grant. Program Number: 4117 Poster Board Number: B0039 Presentation Time: 8:30 AM–10:15 AM Depot Indocyanine Green Dye for in vivo visualization of infiltrating leukocytes Dawn A. Sim1, 2, Colin J. Chu2, Michael B. Powner2, Senthil Selvam2, Adnan Tufail1, 2, Catherine A. Egan1, James W. Bainbridge1, 2, Richard W. Lee3, 1, Andrew D. Dick3, 1, Marcus Fruttiger2. 1NIHR Biomedical Research Center, Moorfields Eye Hospital, London, United Kingdom; 2 UCL Institute of Ophthalmology, London, United Kingdom; 3 Academic unit of Ophthalmology, School of Clinical Sciences, University of Bristol, Bristol, United Kingdom. Purpose: To assess a novel method to label and image myeloid cells infiltrating the mouse retina and choroid in vivo, using a single depot injection of Indocyanine green dye (ICG). Methods: We assess the use of depot ICG for the purpose of in vivo cell labelling on three different ocular models of inflammation and angiogenesis - endotoxin-induced uveitis (EIU), experimental autoimmune uveoretinitis (EAU) and laser-induced choroidal neovascularisation (CNV) model. A near-infrared scanning ophthalmoscope was used for in vivo imaging of the eye and flow cytometry was used on blood and spleen to assess the number and phenotype of labelled cells. ICG was administered 72 hours prior to the induction of inflammation to ensure clearance from systemic circulation. Results: We found that in vivo intravenous administration failed to label any leukocytes, whereas depot injection, either intraperitoneal or subcutaneous, was successful in labelling CD11b+ myeloid cells. Progression of inflammation could be traced over a period of 14 days following a single depot injection of ICG. Additionally, flow cytometric analysis revealed that the predominant population of cells stained by ICG are circulating and splenic reservoir CD11b+ myeloid cells. Commercial Relationships: Dawn A. Sim, Allergan (F), UCL Business (P); Colin J. Chu, None; Michael B. Powner, None; Senthil Selvam, None; Adnan Tufail, UCL Business (P); Catherine A. Egan, None; James W. Bainbridge, None; Richard W. Lee, None; Andrew D. Dick, None; Marcus Fruttiger, UCL Business (P) Program Number: 4118 Poster Board Number: B0040 Presentation Time: 8:30 AM–10:15 AM Exploring photoreceptor directionality using quadrant pupil detection Brian Vohnsen, Salihah Qaysi, Pedro Anjos, Denise Valente. School of Physics, University College Dublin, Dublin, Ireland. Purpose: Photoreceptors are endowed with directional properties that are expressed by their angular sensitivity to incident light and directionality in retinal imaging known as the Stiles-Crawford effect of the first kind (SCE) and the optical SCE, respectively. Individual photoreceptor tilt can be analyzed by moving the incident light across the pupil when capturing retinal images. The purpose of this study is to examine with numerical analysis and fundus imaging the viability of an alternative quadrant detection scheme in which retinal images are captured through four equal-sized pupil sectors from which individual cone photoreceptor tilts can be derived. Methods: A numerical analysis has been performed for directional light scattering from photoreceptor cones in the retinal mosaic modeled as tri-layered structures that represent the location of mitochondria in the ellipsoid, the inner-outer segment junction and the posterior outer segment tip [Vohnsen, Frontiers in Optics (2014) FW5F.5]. Each outer segment has been added random tilt of varying amplitude. Simulated cone mosaic images are calculated through 4 equal-sized sectors in the pupil plane. The numerical predictions are compared with experimental quadrant parafoveal fundus images for healthy subjects. ©2015, Copyright by the Association for Research in Vision and Ophthalmology, Inc., all rights reserved. Go to iovs.org to access the version of record. For permission to reproduce any abstract, contact the ARVO Office at [email protected]. ARVO 2015 Annual Meeting Abstracts Results: The sectored quadrant pupil allows not only determination of total intensity images but also direct determination of local cone photoreceptor tilts (which may differ from their psychophysically relevant tilts) in two orthogonal directions using the sectored backscattered light intensity. It is found that the method is highly suited to determine tilt without requiring displacement of the incident light in the pupil plane whereby the analysis becomes simplified although it requires proper centration with respect to the SCE peak location. Retinal images in the healthy subjects show only small photoreceptor tilts in the parafoveal region but confirm the potential of the technique. Conclusions: The numerical analysis shows that the quadrant pupil detection is suitable for rapid detection of individual photoreceptor tilts in the photoreceptor mosaic of less than 1 degree. With proper optimization this may have clinical potential for analysing perturbations caused by drusen and retinal disease that may alter the pointing of the photoreceptors causing a resulting deterioration of vision. Commercial Relationships: Brian Vohnsen, None; Salihah Qaysi, None; Pedro Anjos, None; Denise Valente, None Support: SFI 08/IN.1/B2053 Program Number: 4119 Poster Board Number: B0041 Presentation Time: 8:30 AM–10:15 AM Intraocular scattering compensation in macular pigment density measurement Dimitrios Christaras, Harilaos Ginis, Alexandros Pennos, Pablo Artal. Laboratorio de Optica, Universidad de Murcia, Murcia, Spain. Purpose: Intraocular scattering reduces the contrast of fundus images, especially in flood-illumination imaging systems. Since macular pigment density (MPD) is often estimated from the analysis of multispectral retinal images, this contrast reduction may affect the results. We performed experiments to better understand the severity of this problem and to overcome it by compensating scatter, thus obtaining a more accurate estimate of the MPD. Methods: A new multispectral fundus imaging system based on the double pass principle, also capable of measuring intraocular scattering at various wavelengths, was built. In addition, a computational method to estimate MPD from retinal images after compensation of light scattering was developed. The experimental setup includes a xenon lamp filtered by band-pass filters and homogenized by diffusers,a motorized iris conjugated to the subject’s retina and an electron multiplying CCD camera. To avoid reflections and backscattered light from the lens and the cornea, the imaging and illumination arms are spatially separated by means of rectangular apertures conjugated to the pupil plane. Intraocular scattering was estimated using an optical integrator method by projection of uniform disks ranging from 0.4 to 6 degrees radius (Ginis, et al (2012), Journal of Vision, 12(3), 1–10). For the MPD calculation, a pair of foveal images was recorded at two wavelengths, 460 and 550 nm. These images were then corrected for the effect of scatter by using a standard deconvolution technique and the scatter free value of MPD was calculated according to a formula (Delori et al (2001), J. Opt. Soc. Am. A, 18, 1212–1230). Results: The complete procedure was successfully applied to 2 young subjects, LH and DC, with no known eye pathology. The observed maximum value of the MPD was 0.34 and 0.28 D.U. for DC and LH respectively, whereas without any scatter compensation the respected vales were 0.18 and 0.16 D.U., exhibiting an underestimation at the order of 45% for the maximum value and 35% for the mean value of the total MPD. The effect depends highly on the overall scattering but also on the difference in scattering for blue and for green. Conclusions: Initial results from two young, healthy subjects have shown an important underestimation of the MPD when the effect of scattering is not taken into account. This effect is expected to be more important in elderly subjects with elevated intraocular straylight. Commercial Relationships: Dimitrios Christaras, None; Harilaos Ginis, None; Alexandros Pennos, None; Pablo Artal, None Program Number: 4120 Poster Board Number: B0042 Presentation Time: 8:30 AM–10:15 AM Methods for an Enhanced 3D Interpretation of Autoradiographic Ocular Drug Distribution Data Julie E. Whitcomb1, Susan S. Lee1, Michael Robinson1, Jie Shen1, Elizabeth Spencer3, Meagan Krueger3, Harvey Pollack2, Rex Moats2, Yang Tang2, Mayssa Attar1. 1Allergan, Irvine, CA; 2Childrens Hospital of Los Angeles, Los Angeles, CA; 3Covance, Madison, WI. Purpose: Understanding drug distribution in the eye following various routes of administration is important for optimizing drug delivery. The physiologic and anatomic barriers that affect drug distribution to the site of action can be studied with autoradiography (ARL), which detects radiolabeled drug compounds on two dimensional film, but lacks anatomical landmarks for spatial reference. Herein, we describe a method to enhance ARL images for 3D interpretation. Methods: A model radiolabeled compound (MW: 430 g/mol) was dosed unilaterally via daily repeated topical dose and a single intravitreal injection (n = 2 monkeys). The frozen heads were sectioned into 43 transverse cross sections 20 mm thick through the eye. Sections were mounted on a lexan plate, photographically imaged (Figure 1a), then exposed on phosphor imaging screens (Figure 1b) with blood standards for radioactivity quantitation. A novel algorithm was designed to align the anatomical photographic images to the ARL slides. Radioactivity concentration as a function of image density was generated and the individual images stacked to render a 3D volume. Results: The 3D reconstruction provided an anatomically accurate and quantitative representation of the 2D phosphor images. The blood standards were used to translate the log-linear concentration of the radioactivity to a quantitative heat map (Figure 1c) to interpretat the dimensional gradient. Interpolation between the individual slices was done to visually illustrate the regional drug distribution between the two routes of administration (Figure 2). Overall the topical dosing resulted in localized distribution in the anterior region; whereas, the intravitreal dose was more dispersed throughout the entire eye. Conclusions: Traditional ARL films are generally difficult to interpret in a 3D space and the drug distribution is difficult to interpret without any anatomic references. Quantitative 3D renderings of drug distribution from 2D digital autoradiographic images is a useful tool for understanding local drug delivery in relation to the anatomical structures and can be applied to other labeling techniques to better understand drug distribution. ©2015, Copyright by the Association for Research in Vision and Ophthalmology, Inc., all rights reserved. Go to iovs.org to access the version of record. For permission to reproduce any abstract, contact the ARVO Office at [email protected]. ARVO 2015 Annual Meeting Abstracts Photographic (top), Autoradiographic (middle), and Quantitative Heat Map (bottom) Images with Topical (left) and Intravitreal (right) Administration 3D Rendered Images Comparing Topical (left) and Intravitreal (right) Administration Commercial Relationships: Julie E. Whitcomb, Allergan (E); Susan S. Lee, Allergan (E); Michael Robinson, Allergan (E); Jie Shen, Allergan (E); Elizabeth Spencer, Covance (E); Meagan Krueger, Covance (E); Harvey Pollack, Allergan (C), Childrens Hospital of Los Angeles (E); Rex Moats, Allergan (C), Childrens Hospital of Los Angeles (E); Yang Tang, Allergan (C), Childrens Hospital of Los Angeles (E); Mayssa Attar, Allergan (E) Program Number: 4121 Poster Board Number: B0043 Presentation Time: 8:30 AM–10:15 AM Microfocus computed tomography as a new method for highresolution imaging of microstructures in the eye: a pilot study Christian Enders1, Eva Braig2, Jens U. Werner1, Gerhard K. Lang1, Gabriele E. Lang1, Kai H. Scherer2, Franz Pfeiffer2. 1Department of Ophthalmology, University of Ulm, Ulm, Germany; 2Department of Pyhsics and Institute of Medical Engineering, Technische Universität München, Munich, Germany. Purpose: Histopathological examination requires processing and provides a two-dimensional (2D) view on the sections of a specimen. The aim of this study is to investigate the structural characteristics of the eye in an ex-vivo study using high-resolution microfocus computed tomography (micro ct) without the need of processing and providing three-dimensional (3D) non-destructive sectional images. Methods: The study was approved by the institutional reviewboard, and informed consent was obtained. A total of 4 specimens were used. Among them were 3 enucleated human bulbi and 1 exenterated swine eye. The 3 human specimens had underwent regular histopathological examination. Diagnoses of the human specimens were angle closure glaucoma, retinal detachment and malignant melanoma. The swine eye had been used for surgical training and a CyPass micro-stent® had been implanted. All specimens were fixated in paraffin. We obtained images with a high-resolution micro ct system (v|tome|x by GE®) for 3D computed tomography and 2D non-destructive X-ray inspection. The maximum resolution is 2-20 micrometers depending on the sample size. Results: The obtained images showed good correlation with histopathologic findings. The maximum resolution in our specimens is 20 micrometers. In the 2D-images retinal, choroidal, uveal, corneal and lental tissues can be distinguished among others. The configuration of the anatomical structures e.g. the chamber-angle can be assessed without damaging the integrity of the specimen. The exact position of a surgically applicated device like the CyPass micro-stent® can be assessed faster and more accurate than with standard histological examination. The 3D-images give a helpful overview of the anatomical structures but cannot distinguish between tissues as good as the 2D-images. Conclusions: The results of this study indicate that ex-vivo micro ct is a helpful research tool in the examination of enucleated bulbi with excellent correlation to histo-pathologic findings. Although at present not applicable in vivo because of the radiation load and sample size, micro ct is a valuable complimentary tool in ex-vivo diagnostics and in the development of new surgical techniques and ophthalmic devices. 3D rendering of a swine eye with a micro-stent orthogonal 2D slice of a a swine eye with a micro-stent Commercial Relationships: Christian Enders, None; Eva Braig, None; Jens U. Werner, None; Gerhard K. Lang, None; Gabriele E. Lang, None; Kai H. Scherer, None; Franz Pfeiffer, None ©2015, Copyright by the Association for Research in Vision and Ophthalmology, Inc., all rights reserved. Go to iovs.org to access the version of record. For permission to reproduce any abstract, contact the ARVO Office at [email protected]. ARVO 2015 Annual Meeting Abstracts Program Number: 4122 Poster Board Number: B0044 Presentation Time: 8:30 AM–10:15 AM Two-photon Signals From Label-free Human Epiretinal Membranes Juan M. Bueno1, Xavier Valldeperas2, Francisco J. Avila1, Raquel Palacios1, Pablo Artal1. 1Laboratorio de Optica, Universidad de Murcia, Murcia, Spain; 2Hospital Universitari Germans Trias, Badalona, Spain. Purpose: Epiretinal membranes (ERMs) are composed of neural elements and different cellular and extracellular matrix (ECM) components usually visualized using multi-labeling methods. We have used two-photon microscopy to image non-stained human ERMs to analyze the sources of two-photon excitation fluorescence (TPEF) and second harmonic generation signal (SHG). This would help to better understand the organization and structure of ERMs. Methods: A custom-developed multiphoton microscope equipped with adaptive optics (Bueno et al., J. Biomed. Opt. 2010) was used to record pairs of TPEF-SHG images originated from the ERM structures in a backward configuration. Specimens from ten patients were successfully removed during vitrectomy and ERM peeling, without the use of ophthalmic dyes. Immediately after surgery, ERMs were fixated with a paraformaldehyde solution overnight and flatmounted for two-photon microscopy imaging. Series of images across the specimen and 3D stacks as a function of depth were acquired for the different samples. Results: Both TPEF and SHG signals were obtained from the ERM’s samples, revealing different structures. SHG signals confirmed the presence of filamentous structures corresponding to collagen fibers and covering the specimen. TPEF signal was significantly higher probably due to the presence of numerous types of cells (Muller, glial…). This cellular autofluorescence is originated from the mitochondrial reduced pyridine nucleotides NAD(P)H and flavin compounds. Moreover, the elastin of the ECM might also contribute to TPEF images. Conclusions: Two-photon microscopy provides a non-invasive tool to image unstained isolated ERMs. TPEF/SHG signal combination revealed detailed information on the ERM morphology not available with classical imaging techniques. Since staining procedures are not required, the sample manipulation is reduced. Non-linear imaging techniques are useful in the visualization and analysis of healthy and pathological retinal and epiretinal tissues. Farther studies should investigate the dependence between TPEF/SHG signals and ERM etiology (idiopathic, diabetic or inflammatory). Commercial Relationships: Juan M. Bueno, None; Xavier Valldeperas, None; Francisco J. Avila, None; Raquel Palacios, None; Pablo Artal, None Support: Ministerio de Economía y Competitividad, Spain (grant FIS2013-41237-R) Program Number: 4123 Poster Board Number: B0045 Presentation Time: 8:30 AM–10:15 AM Noninvasive functional retinal imaging disrupted by pharmacologic manipulation of neurovascular coupling pathways Daniel Y. Tso1, 2, Momotaz Begum1, 2. 1Depts of Neurosurgery and Ophthalmology, SUNY Upstate Medical Univ, Syracuse, NY; 2SUNY Eye Institute, Syracuse, NY. Purpose: Intrinsic signal optical imaging studies have shown the existence of stimulus-evoked signals in the retina that are likely to be of outer retinal origin. Additional studies have also shown that such reflectance change signals are dominated by hemodynamics. The linkage between this stimulus-driven retinal activity and the resulting hemodynamic basis of the measured reflectance changes is a form of neurovascular coupling. However the precise chain of mechanisms in the retina whereby photoreceptor activation leads to a vascular response is unknown. Most previous studies of neurovascular coupling in the retina have been in vitro. In the present study, we sought to dissect the role of several prospective signaling pathways in the observed stimulus-evoked neurovascular responses in vivo through intravitreal injections of selected agonists, antagonists and blockers, while imaging these retinal intrinsic signals. Methods: Animals and Preparation. Adult cats were anesthetized, paralyzed and positioned in a stereotaxic. Using a modified fundus camera, the retina was stimulated with visible (550nm) patterned stimuli and illuminated in the near-infrared (700-900nm), while intrinsic optical signals were recorded with a CCD camera. Drug-injections. Intravitreal injections included suramin (purinergic antagonist), HET0016 (20-HETE inhibitor), PPOH (EET inhibitor) and NECA (adenosine agonist). Results: Previous retinal imaging studies using intravitreal injections of blockers of inner retinal function (e.g. TTX, PDA, APB) yielded little impact on the retinal imaging signals. However in the present study, each drug selected to interfere with a particular neurovascular coupling pathway had profound impact on the observed retinal imaging signals. In some cases, the sign of the observed retinal imaging signal changed from negative to positive. Conclusions: This novel functional retinal imaging technique is shown to reveal outer retinal function through its impact on hemodynamics via the neurovascular coupling pathways. These studies help establish the chain of retinal events from light absorption to observed changes in retinal reflectance in vivo. It is anticipated that an understanding of the underlying neural, biophysical and signaling mechanisms will help determine the utility of this method for studies of retinal function, health and related clinical applications. Commercial Relationships: Daniel Y. Tso, None; Momotaz Begum, None Program Number: 4124 Poster Board Number: B0046 Presentation Time: 8:30 AM–10:15 AM RetinAsk: A Queryable Atlas of the Retina Jessica Taibl, Samir I. Sayegh. The EYE Center, Champaign, IL. Purpose: To present the improved version of The Queryable Atlas of the Retina that allows for queries based on partial information, including retinal images, of an unknown retinal condition. Methods: Since the original presentation of the Queryable Atlas of the Retina (Sayegh et al, AAO 2000) a number of events have occurred that led to a fundamental redesign and improvement of the Atlas. 1) OCT has become a major tool of diagnosis of retinal disease, 2) the speed and memory capacity of computers, as well as speed of implementation of search algorithms, have continued to rise exponentially and 3) new treatments paradigm and novel surgical techniques have profoundly transformed retinal therapeutics. These three aspects have been utilized to redesign the Atlas while ©2015, Copyright by the Association for Research in Vision and Ophthalmology, Inc., all rights reserved. Go to iovs.org to access the version of record. For permission to reproduce any abstract, contact the ARVO Office at [email protected]. ARVO 2015 Annual Meeting Abstracts maintaining its fundamental queryable nature which clearly distinguishes it from other more traditional approaches. Results: The new design allows for rapid queries to be resolved based on limited characterization or partial information. The search “by field” and “by image” features have been vastly improved. Conclusions: The Queryable Atlas of the Retina can be used as a powerful diagnostic support tool as well as a valuable teaching tool in residency and retinal fellowship programs. Commercial Relationships: Jessica Taibl, None; Samir I. Sayegh, None Program Number: 4125 Poster Board Number: B0047 Presentation Time: 8:30 AM–10:15 AM Physical and Optical Properties of Rigid Gas Permeable Contact Lenses Measured with Optical Coherence Tomography Dean A. VanNasdale, Gang Huang, Nicky Lai, Marisa Ciamacca, Amanda Eilerman, Aaron Zimmerman, Alex Nixon, Molly Smith. Optometry, Ohio State Univ College of Optometry, Columbus, OH. Purpose: To calibrate the axial and lateral dimensions of spectral domain optical coherence tomography (SDOCT) images using rigid gas permeable (RGP) contact lenses. To quantify the physical and optical characteristics of RGP lenses using cross sectional OCT imaging. Methods: The parameters of 5 spherical RGP lenses of the same design, index of refraction = 1.433, were manually verified. The posterior side of each lens was mounted on a flat surface and imaged using the Heidelberg Spectralis SDOCT system with Anterior Segment Module (Heidelberg Engineering, Carlsbad, CA). The 30 deg image included the entire cross section of the central lens and extended beyond the lens boundaries to included locations of the mounting platform only, which was used as an axial reference. The lateral and axial pixel dimensions were calculated and divided by the known diameter of the contact lens and center thickness. The index of refraction of the lens material was used to compensate for the thickness changes induced in the cross sectional imaging. Optical path length differences created by the thickness and index of refraction change of the lenses were confirmed by measuring by the displacement of the flat mounting surface under the lens, relative to the undeviated surface imaged outside of the lens boundaries. Results: Lateral and axial characteristics of the OCT images could be successfully calibrated using the known parameters of the RGP lenses. The coefficient of variation for distance conversions of the 5 lenses was less than 1% for each of the axial and lateral dimensions. The displacement of the flat surface caused by the optical path length difference induced by the contact lens was linearly proportional to the thickness of the contact lens (r2 = 0.9736). Conclusions: RGP lenses can be used to calibrate cross sectional OCT images in both the lateral and axial dimensions. Physical characteristics of RGP lenses can be extracted from cross sectional OCT data. Optical properties of RGP lens can be assessed using optical path length differences induced by the RGP thickness and index of refraction changes. Commercial Relationships: Dean A. VanNasdale, None; Gang Huang, None; Nicky Lai, None; Marisa Ciamacca, None; Amanda Eilerman, None; Aaron Zimmerman, None; Alex Nixon, None; Molly Smith, None ©2015, Copyright by the Association for Research in Vision and Ophthalmology, Inc., all rights reserved. Go to iovs.org to access the version of record. For permission to reproduce any abstract, contact the ARVO Office at [email protected].