Multimodal imaging of cryofixed cells using Raman and fluorescence structured illumination microscopy

Kosuke Tsuji; Masahito Yamanaka; Yasuaki Kumamoto; Shoko Tamura; Kenta Mizushima; Kakeru Kono; Toshiki Kubo; Hanae Hirano; Kazunori Sugiura; Shun-Ichi Fukushima; Takumi Kunimoto; Kentaro Nishida; Yoshinori Harada; Nicholas I. Smith; Rainer Heintzmann; Takeharu Nagai; Hideo Tanaka; Katsumasa Fujita

doi:10.1117/12.3000361

13 March 2024 Multimodal imaging of cryofixed cells using Raman and fluorescence structured illumination microscopy

Kosuke Tsuji, Masahito Yamanaka, Yasuaki Kumamoto, Shoko Tamura, Kenta Mizushima, Kakeru Kono, Toshiki Kubo, Hanae Hirano, Kazunori Sugiura, Shun-Ichi Fukushima, Takumi Kunimoto, Kentaro Nishida, Yoshinori Harada, Nicholas I. Smith, Rainer Heintzmann, Takeharu Nagai, Hideo Tanaka, Katsumasa Fujita

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Proceedings Volume PC12846, Imaging, Manipulation, and Analysis of Biomolecules, Cells, and Tissues XXII; PC128460B (2024) https://doi.org/10.1117/12.3000361
Event: SPIE BiOS, 2024, San Francisco, California, United States

Abstract

Using complementary optical microscopy techniques provides more detailed insight into biological samples. However, misinterpretation can occur by temporal discrepancies due to differences in temporal resolution and switching imaging modalities. Here, we demonstrated multimodal imaging of cryofixed cells using Raman and fluorescence structured illumination microscopy (SIM). Cryofixation preserves structures and chemical states of samples in their near-native states, allowing multimodal imaging without artifacts caused by temporal discrepancy. We demonstrated multimodal imaging of cryofixed HeLa cells stained with an actin probe, where Raman microscope visualized cytochromes, proteins and lipids, and SIM visualized fluorescence-labelled actin filaments.

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Kosuke Tsuji, Masahito Yamanaka, Yasuaki Kumamoto, Shoko Tamura, Kenta Mizushima, Kakeru Kono, Toshiki Kubo, Hanae Hirano, Kazunori Sugiura, Shun-Ichi Fukushima, Takumi Kunimoto, Kentaro Nishida, Yoshinori Harada, Nicholas I. Smith, Rainer Heintzmann, Takeharu Nagai, Hideo Tanaka, and Katsumasa Fujita "Multimodal imaging of cryofixed cells using Raman and fluorescence structured illumination microscopy", Proc. SPIE PC12846, Imaging, Manipulation, and Analysis of Biomolecules, Cells, and Tissues XXII, PC128460B (13 March 2024); https://doi.org/10.1117/12.3000361

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KEYWORDS

Raman spectroscopy

Multimodal imaging

Biological imaging

Fluorescence

Light sources and illumination

Optical microscopy

Biological samples

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