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.
Yuta Suzuki, Koya Kobayashi, Dinghuan Deng, Yoshifumi Wakisaka, Keisuke Goda, and Yasuyuki Ozeki, "Multicolor stimulated Raman imaging of live microalgal cells using fast wavelength-switched laser pulses (Conference Presentation)," Proc. SPIE 10505, High-Speed Biomedical Imaging and Spectroscopy III: Toward Big Data Instrumentation and Management, 105050N (Presented at SPIE BiOS: January 30, 2018; Published: 15 March 2018); https://doi.org/10.1117/12.2291189.5751641653001.
Conference Presentations are recordings of oral presentations given at SPIE conferences and published as part of the conference proceedings. They include the speaker's narration along with a video recording of the presentation slides and animations. Many conference presentations also include full-text papers. Search and browse our growing collection of more than 12,000 conference presentations, including many plenary and keynote presentations.