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PreprintCited by 2Digital Holography and MicroscopyAdvanced Fluorescence Microscopy TechniquesCell Image Analysis Techniques

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Label-free imaging of intracellular structures in living mammalian cells via external apodization phase-contrast microscopy

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Ohno, Hiroshi, Nishimura, Takenori, Kainoh, Kenta, Ohashi, Yoshitaka, Onodera, Naoko, Kano, Mayuko, Sari, Lay Nurhana, Masuda, Masato, Tamura, Yoshiaki, Hayashi, Yusuke, Yamamoto, Yusuke, Takahashi, Shin-Ichiro, Mishima, Yuta, Yoneyama, Yosuke, Takeuchi, Yoshinori, Sekiya, Motohiro, Matsuzaka, Takashi, Miyamoto, Takafumi, Shimano, Hitoshi·2024·FEBS Journal
DOI 10.1101/2024.03.01.582671PMC PMC12914761

Preprint — published as 10.1111/febs.70286 · Paper page

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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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Metadata

Authors
Ohno, H., Nishimura, T., Kainoh, K., Ohashi, Y., Onodera, N., Kano, M., Sari, L. N., Masuda, M., Tamura, Y., Hayashi, Y., Yamamoto, Y., Takahashi, S.-I., Mishima, Y., Yoneyama, Y., Takeuchi, Y., Sekiya, M., Matsuzaka, T., Miyamoto, T., & Shimano, H.
Affiliations
  • University of Tsukuba (JP)
  • The University of Tokyo (JP)
  • Toyo University (JP)
  • National Cancer Centre Japan (JP)
  • Tokyo Medical and Dental University (JP)
Corresponding author
Miyamoto, Takafumi
ORCID
Venue
FEBS Journal
Publisher
openRxiv
Published
2024-03-02
Volume
293
Issue
4
Pages
1127-1149
ISSN
1742-464X (ISSN-L)
DOI
10.1101/2024.03.01.582671
PMC
PMC12914761
OpenAlex
W4415286773
Cited by
2
References
74
Citations by year
2026: 1 · 2025: 1
FWCI
0.73
Type
journal article
Language
en
Open access
Licence
Locations
  • FEBS Journal (publishedVersion, cc-by)
  • bioRxiv (Cold Spring Harbor Laboratory) (acceptedVersion, cc-by)
  • PubMed (publishedVersion)
  • PubMed Central (publishedVersion, cc-by)
Funders
  • Takeda Science Foundation (DGM06005J)
  • Cabinet Office, Government of Japan (#JPJ009237)
  • University of Tsukuba (JPMJPF2017)
  • Nature
  • Japan Society for the Promotion of Science (JPMJPF2017, 24K14766, 23K23562, KAKENHI (22H02296), CREST)
  • Japan Science and Technology Agency (JPMJPF2017)
  • Bio-oriented Technology Research Advancement Institution (JP J009237)
  • Core Research for Evolutional Science and Technology (JPMJCR1927)
  • Co-creation place formation support program (JPMJPF2017)
  • Moonshot Research and Development Program (JPJ009237)
Topics
  • Physical Sciences › Physics and Astronomy › Atomic and Molecular Physics, and Optics › Digital Holography and Microscopy
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Biophysics › Advanced Fluorescence Microscopy Techniques
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Biophysics › Cell Image Analysis Techniques
Keywords
label-free imaging, intracellular structures, biomolecular condensates, lipid droplets, mitochondria, live-cell imaging, spatiotemporal resolution
MeSH terms
Biomolecular Condensates, Animals, HeLa Cells, Humans, Microscopy, Fluorescence, Microscopy, Phase-Contrast, Mitochondria, Lipid Droplets

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