Download Methods Discussion Abstract Graduate

Graduate
Category: Engineering and Technology
Degree Level: PhD
Abstract ID#: 80
David Walsh, Juliette Kassas, Shashi K. Murthy
Abstract
Improper diagnosis of idiopathic eye disease can have disastrous
Methods
Discussion
Current approach is limited in efficacy due to: 1) Low number of cells and 2) Fragility of sample
Research impacts:
Solution: A label-free separation method that can be performed at the point-of-care (POC)
results. Uveitis and primary intraocular lymphoma (PIOL) are
• Patient can get diagnosis during visit
• Save money on materials, equipment, and lab tests
symptomatically similar diseases but require drastically different
treatments. Current methods have a diagnostic yield of only 20% [1].
1. Collect patient sample via pars plana vitrectomy.
• Small footprint and portable (7.5 cm x 2.5 cm x 0.3 cm)
We are developing a point-of-care platform to rapidly characterize cells
2. Establish a chemotactic gradient and place cells on the testing
• No need for experienced lab technicians
within the eye for definitive diagnosis. This method will minimize cost
• Preserves sample for further analysis
platform.
and time, while improving fidelity of results. The implementation of
• Prevents scarring of eye tissue and permanent vision loss
the system at the point-of-care will ensure proper treatment.
• Determines accurate course of treatment
Background
SDF-1α
BCA-1
• ~2.3 million Americans suffer from uveitis, and vision-loss due to
uveitis accounts for 10-15% of the blindness in the U.S. [2].
• Primary intraocular lymphoma (PIOL) has a two-year survival rate
of 39% [3, 4].
Device prototype
• PIOL is commonly referred to as masquerade syndrome due to its
gradual onset and ability to mimic other ocular diseases [5].
• The current diagnostic approach analyzes a biopsy sample from the
Conclusion
vitreous humor.
• A definitive diagnosis is achieved by characterizing cells within the
eye; an invasion of T-lymphocytes implicates uveitis while B-
Attracts B
lymphocytes
(PIOL)
Attracts T
lymphocytes
(Uveitis)
lymphocytes implicate PIOL.
• There is an urgent need to fill the technological gap for diagnosis of
T- and B-lymphocytes within the vitreous biopsy are too similar to
be separated solely on physical properties. Chemotaxis provides a
label-free separation alternative. We have started the development of
a paper-based, chemotactic, cell immunophenotyping device. This
idiopathic eye disease.
process bypasses cell transit from the clinic to the lab and additional
preprocessing steps, which can frequently impact cell integrity and
viability, compromising analysis. Implementation at the point-of-
4. Document the migration path of each cell over a 20 minute period,
care will rapidly diagnose uveitis and PIOL, improving overall
using CellProfiler software.
patient prognoses.
5. Calculate the chemotactic index (C.I.) as displacement towards
*CellProfiler was developed by the Broad Institute
gradient (x) over total migration distance (d).
6. Determine if C.I. value indicates if cellular movement is due to
implementation of the gradient.
Current diagnostic approaches involve analysis by flow cytometry and histology.
Acknowledgements:
This material is based upon work supported by the National Science Graduate Research
Fellowship awarded to D.I.W. under grant number NSF/DGE-0946746. Any opinion, findings
and conclusions or recommendations expressed in this material of those of the author(s) and do
not necessarily reflect the views of the National Science Foundation
PIOL is also known as
masquerade syndrome.
References:
1. Margolis, R., et al., Vitrectomy for the diagnosis and management of uveitis of unknown cause. Ophthalmology, 2007. 114(10): p. 1893-7.
2. Durrani, O.M., C.A. Meads, and P.I. Murray, Uveitis: a potentially blinding disease. Ophthalmologica, 2004. 218(4): p. 223-36.
3. Ferreri, A.J., et al., Relevance of intraocular involvement in the management of primary central nervous system lymphomas. Ann Oncol, 2002. 13(4): p. 531-8.
4. Schabet, M., Epidemiology of primary CNS lymphoma. J Neurooncol, 1999. 43(3): p. 199-201.
5. Korfel, A., et al., Das Masquerade-Syndrom. Dtsch Arztebl, 2007. 104(8): p. 490-5.
*Lin, F., et al., Lab Chip. 2006.
x
C. I. 
d