Label‐free imaging of intracellular structures in living mammalian cells via external apodization phase‐contrast microscopy
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 apodized 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 previously unknown cellular mechanisms.
Authors
- Yusuke Hayashi (ORCID: https://orcid.org/0000-0001-5672-1497)
- Motohiro Sekiya (ORCID: https://orcid.org/0000-0002-5170-9699)
- Yuta Mishima
- Kenta Kainoh
- Hitoshi Shimano (ORCID: https://orcid.org/0000-0002-5562-5572)
- Masato Masuda
- Yosuke Yoneyama (ORCID: https://orcid.org/0000-0002-9170-2846)
- Takashi Matsuzaka (ORCID: https://orcid.org/0000-0002-5898-3463)
- Yusuke Yamamoto (ORCID: https://orcid.org/0000-0002-5262-8479)
- Takafumi Miyamoto (ORCID: https://orcid.org/0000-0001-9363-2864)
- Takenori Nishimura
- Hiroshi Ohno (ORCID: https://orcid.org/0000-0001-8776-9661)
- Teppei Nishino (ORCID: https://orcid.org/0000-0003-2544-4639)
- Yoshinori Takeuchi (ORCID: https://orcid.org/0000-0003-2717-5719)
- Shin‐Ichiro Takahashi (ORCID: https://orcid.org/0000-0002-2323-2010)
- Yoshiaki Tamura
- Yoshitaka Ohashi
- N. Onodera
- Mayuko Kano
- Lay Nurhana Sari
Institutions
- Tokyo Medical and Dental University (JP)
- University of Tsukuba (JP)
- National Cancer Centre Japan (JP)
- Laboratoire d’immunologie intégrative du cancer (FR)
- The University of Tokyo (JP)
- University of Human Arts and Sciences (JP)
Publication Details
- Journal
- FEBS Journal
- Published
- 2025-10-17
- DOI
- https://doi.org/10.1111/febs.70286
- Primary Topic
- Heat shock proteins research
- Type
- article
- Field-Weighted Citation Impact
- 0.00
Funders
- Cabinet Office, Government of Japan
- University of Tsukuba
- Nature
- Japan Society for the Promotion of Science
- Bio-oriented Technology Research Advancement Institution
- Core Research for Evolutional Science and Technology
- Co-creation place formation support program
- Moonshot Research and Development Program