High-sensitivity Raman microscopy of cryofixed biological samples

Katsumasa Fujita; Kenta Mizushima; Shoko Tamura; Masahito Yamanaka; Menglu Li; Yoshinori Harada; Nicholas I. Smith; Yasuaki Kumamoto; Hideo Tanaka

doi:10.1117/12.3000955

13 March 2024 High-sensitivity Raman microscopy of cryofixed biological samples

Katsumasa Fujita, Kenta Mizushima, Shoko Tamura, Masahito Yamanaka, Menglu Li, Yoshinori Harada, Nicholas I. Smith, Yasuaki Kumamoto, Hideo Tanaka

Author Affiliations +

Proceedings Volume PC12839, Biomedical Vibrational Spectroscopy 2024: Advances in Research and Industry; PC1283908 (2024) https://doi.org/10.1117/12.3000955
Event: SPIE BiOS, 2024, San Francisco, California, United States

Abstract

Raman microscopy provides a variety of insights into molecular composition, chemical state, and environmental conditions in biological samples. However, biological imaging with Raman microscopy have faced challenges such a low signal-to-noise ratio, mainly due to the low scattering efficiency of Raman scattering. To overcome this limitation, we developed a cryo-Raman microscope integrated with a cryostat capable of rapid freezing of biological samples and low-temperature Raman imaging. The spatiotemporal cryofixation of biological samples allows long exposure measurements to accumulate signals without photodamage. We observed both reduction of photobleaching in resonant Raman scattering of cytochromes in cryofixed HeLa cells, and the preservation of redox states of cytochromes in rat heart tissue by cryofixation.

Conference Presentation

Citation Download Citation

Katsumasa Fujita, Kenta Mizushima, Shoko Tamura, Masahito Yamanaka, Menglu Li, Yoshinori Harada, Nicholas I. Smith, Yasuaki Kumamoto, and Hideo Tanaka "High-sensitivity Raman microscopy of cryofixed biological samples", Proc. SPIE PC12839, Biomedical Vibrational Spectroscopy 2024: Advances in Research and Industry, PC1283908 (13 March 2024); https://doi.org/10.1117/12.3000955

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