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Construction of Confocal Raman Microspectrometer and Examination of Possible Modifications for the Detection of Bacteria in vivo Shannon Harrison Megan McInerney Advisor: Anita Mahadevan-Jansen Project Description • Immediate Goals: – Construct Confocal Raman Microspectometer similar to one designed by G.J. Puppels – Propose design modification for use on living subjects – Create spreadsheet incorporating spectral data – Design molecular bond database • Ultimate Desire: Construct a probe utilizing confocal Raman microspectroscopy that will accurately detect and identify bacteria present in middle ear causing infection Motivation and Benefits • Eliminate “troubleshooting” approach to treating middle ear infections • Construct a confocal Raman system that can be used for in vivo analysis • Reduce cost of health care • Proposed probe design more sterile, less expensive and quicker than swab analysis Raman Spectroscopy • What • Why – Advantages – Disadvantages • How System Requirements • Short integration time • Input spectral data • Analyze the spectra by matching the peaks to a database of standards • Output responsible bacteria • Portable Confocal Raman Microspectometer Design Flowchart Apparatus Modifications • Using fiber optic cables, construct probe including the objective and bandpass filters • Align and fix optical components to make apparatus portable Data Analysis • Understand the basis of Raman Spectroscopy • Compile a list of common spectral peaks • Create a temporary spreadsheet containing the bond and Raman shift • Become familiar with Matlab • Design a program that will input sample spectra and identify the bonds by matching peaks • Use the probe to determine spectra for bacteria causing Otitis Media • Integrate the above spectra into the database program • Test the program for recognition of the bacterial spectra Database Flow Chart Example Spectrum Raman Spectroscopic Information Safety Precautions • Environment • Thermal tissue damage – standard regulation • Maintaining sterile techniques • Accuracy Preliminary Conclusions • Construction, alignment, and preliminary testing feasible by end of semester • Probe designed by end of semester • We suggest that this project is continued next semester by BME 272/273 students to complete construction and testing of probe as well as implement molecular bond database References • Http://www.badertech.com • Http://www.apptechsys.com • Customer service personnel from: – MSDSpro – Nousoft – Ovid • Pedro Pedroso • Anita Mahadevan-Jansen • Introductory Raman Spectroscopy, Ferraro Makamato • Raman Spectroscopy, Long, D.A. • Raman Spectroscopy for the Detection of Cancers and Precancers, Mahadevan-Jansen, Antia • Confocal Raman Spectroscopy, Puppels, G.J.