Label‐free imaging of intracellular structures in living mammalian cells via external apodization phase‐contrast microscopy
Hiroshi Ohnoa b, Takenori Nishimurac d, Kenta Kainoha b, Yoshitaka Ohashia b, Naoko Onoderaa b, Mayuko Kanoa b, Lay Nurhana Saria b, Masato Masudae f, Yoshiaki Tamurae f, Teppei Nishinoa b, Yusuke Hayashig h, Yusuke Yamamotog 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 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.
TL;DRIt is demonstrated 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. (Semantic Scholar)