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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
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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.
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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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KEYWORDS
Raman spectroscopy
Biological imaging
Signal to noise ratio
Biological samples
Cryostats
Microscopy
Raman scattering
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