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Proceedings Paper

Multicolor stimulated Raman imaging of live microalgal cells using fast wavelength-switched laser pulses (Conference Presentation)

Paper Abstract

High-speed label-free imaging with chemical contrast is effective for non-invasive analysis of the metabolic heterogeneity of single cells. Stimulated Raman scattering (SRS) microscopy enables high-speed label-free image acquisition with molecular vibrational specificity. While single-color SRS microscopy only acquires images at a certain vibrational frequency, multicolor SRS microscopy successively acquires SRS images at different vibrational frequencies, which then can be used to investigate the distributions of different intracellular molecules. However, its imaging speed remains an order of magnitude slower than that of single-color video-rate SRS microscopy. Previous approaches to circumvent this issue used either only two colors with limited chemical specificity or multiplex detection of SRS spectra using a photodiode array at the expense of imaging speed. Here we demonstrate high-speed four-color SRS imaging using a single photodiode by introducing fast wavelength-switched laser pulses. The fast wavelength switching is realized by the use of an optical intensity modulator as a time gate, a diffraction grating, and fiber delay lines. Using the developed system, we demonstrate motion-artifact-free multicolor SRS imaging of polymer beads and living cells. The results firmly support that our method is a powerful tool for the label-free analysis of living cells in microbiology, oncology, plant science, and medicine.

Paper Details

Date Published: 15 March 2018
Proc. SPIE 10505, High-Speed Biomedical Imaging and Spectroscopy III: Toward Big Data Instrumentation and Management, 105050N (15 March 2018); doi: 10.1117/12.2291189
Show Author Affiliations
Yuta Suzuki, The Univ. of Tokyo (Japan)
Koya Kobayashi, The Univ. of Tokyo (Japan)
Dinghuan Deng, The Univ. of Tokyo (Japan)
Yoshifumi Wakisaka, The Univ. of Tokyo (Japan)
Keisuke Goda, The Univ. of Tokyo (Japan)
Yasuyuki Ozeki, The Univ. of Tokyo (Japan)

Published in SPIE Proceedings Vol. 10505:
High-Speed Biomedical Imaging and Spectroscopy III: Toward Big Data Instrumentation and Management
Kevin K. Tsia; Keisuke Goda, Editor(s)

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