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 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.