22 March 2013 Raman microspectrometer combined with scattering microscopy and lensless imaging for bacteria identification
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Abstract
In this paper, we report on a compact prototype capable both of lensfree imaging, Raman spectrometry and scattering microscopy from bacteria samples. This instrument allows high-throughput real-time characterization without the need of markers, making it potentially suitable to field label-free biomedical and environmental applications. Samples are illuminated from above with a focused-collimated 532nm laser beam and can be x-y-z scanned. The bacteria detection is based on emerging lensfree imaging technology able to localize cells of interest over a large field-of-view of 24mm2. Raman signal and scattered light are then collected by separate measurement arms simultaneously. In the first arm the emission light is fed by a fiber into a prototype spectrometer, developed by Tornado Spectral System based on Tornado’s High Throughput Virtual Slit (HTVS) novel technology. The enhanced light throughput in the spectral region of interest (500-1800 cm-1) reduces Raman acquisition time down to few seconds, thus facilitating experimental protocols and avoiding the bacteria deterioration induced by laser thermal heating. Scattered light impinging in the second arm is collected onto a charge-coupled-device. The reconstructed image allows studying the single bacteria diffraction pattern and their specific structural features. The characterization and identification of different bacteria have been performed to validate and optimize the acquisition system and the component setup. The results obtained demonstrate the benefits of these three techniques combination by providing the precise bacteria localization, their chemical composition and a morphology description. The procedure for a rapid identification of particular pathogen bacteria in a sample is illustrated.
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S. A. Strola, S. A. Strola, E. Schultz, E. Schultz, C. P. Allier, C. P. Allier, B. DesRoches, B. DesRoches, J. Lemmonier, J. Lemmonier, J.-M. Dinten, J.-M. Dinten, } "Raman microspectrometer combined with scattering microscopy and lensless imaging for bacteria identification", Proc. SPIE 8572, Advanced Biomedical and Clinical Diagnostic Systems XI, 85720X (22 March 2013); doi: 10.1117/12.2002301; https://doi.org/10.1117/12.2002301
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