Label-free imaging of cellular organization in living mammalian cells via external apodization phase-contrast microscopy

Sari, L. N.; Miyamoto, T.; Nishimura, T.; Kainoh, K.; Onodera, N.; Kano, M.; Masuda, M.; Tamura, Y.; Hayashi, Y.; Yamamoto, Y.; Takahashi, S.-I.; Mishima, Y.; Yoneyama, Y.; Takeuchi, Y.; Ohno, H.; Ohashi, Y.; Sekiya, M.; Matsuzaka, T.; Shimano, H.

doi:10.1101/2024.03.01.582671

bioRxiv · 10.1101/2024.03.01.582671

Label-free imaging of cellular organization in living mammalian cells via external apodization phase-contrast microscopy

Sari, L. N.·Miyamoto, T.·Nishimura, T.·Kainoh, K.·Onodera, N.·Kano, M.·Masuda, M.·Tamura, Y.·Hayashi, Y.·Yamamoto, Y.·Takahashi, S.-I.·Mishima, Y.·Yoneyama, Y.·Takeuchi, Y.·Ohno, H.·Ohashi, Y.·Sekiya, M.·Matsuzaka, T.·Shimano, H.

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Abstract

Developing techniques to visualize intracellular structures, which influence the spatiotemporal functionality of biomolecules, is essential for elucidating mechanisms governing cellular behavior. In this study, we demonstrate that label-free external apodization phase-contrast (ExAPC) microscopy serves as a valuable tool for the simultaneous observation of various intracellular structures with high spatiotemporal resolution, while successfully mitigating halo artifacts. Additionally, through quantitative analysis of images obtained by combining ExAPC microscopy with fluorescence microscopy, we identified distinct heterogeneities in biomolecular condensates, lipid droplets, and mitochondria. Our findings highlight the potential of ExAPC microscopy to provide detailed insights into alterations in intracellular structures associated with diverse cellular processes, corroborating the existing knowledge and potentially contributing to the discovery of novel cellular mechanisms.

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Sari, L. N., Miyamoto, T., Nishimura, T., Kainoh, K., Onodera, N., Kano, M., Masuda, M., Tamura, Y., Hayashi, Y., Yamamoto, Y., Takahashi, S.-I., Mishima, Y., Yoneyama, Y., Takeuchi, Y., Ohno, H., Ohashi, Y., Sekiya, M., Matsuzaka, T., Shimano, H.. 2024-03-02. Label-free imaging of cellular organization in living mammalian cells via external apodization phase-contrast microscopy. https://doi.org/10.1101/2024.03.01.582671

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