Holotomographic microscopy reveals label-free quantitative dynamics of endothelial cells during endothelialization

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Abstract

Holotomograhic microscopy (HTM) has emerged as a non-invasive imaging technique that offers high-resolution, quantitative 3D imaging of biological samples. This study explores the application of HTM in examining endothelial cells (ECs). HTM overcomes the limitations of traditional microscopy methods in capturing the real-time dynamics of ECs by leveraging the refractive index (RI) to map 3D distributions label-free. This work demonstrates the utility of HTM in visualizing key cellular processes during endothelialization, wherein ECs anchor, adhere, migrate, and proliferate. Leveraging the high resolution and quantitative power of HTM, we show that lipid droplets and mitochondria are readily visualized, enabling more comprehensive studies on their respective roles during endothelialization. The study highlights how HTM can uncover novel insights into EC behavior, offering potential applications in medical diagnostics and research, particularly in developing treatments for cardiovascular diseases. This advanced imaging technique not only enhances our understanding of EC biology but also presents a significant step forward in the study of cardiovascular diseases, providing a robust platform for future research and therapeutic development.

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  1. Large-field tiled acquisitions highlight nuclei and clear cellboundaries, enabling the simultaneous observation of numerous cells.

    how does this compare to just using standard brightfield imaging or other label-free techniques (e.g. DIC)? are there any features that quantitative phase picks up that other techniques miss?

  2. The precipitous drop in RI valuesmay be attributed to changes in cell density and volume as cells spread out,

    Is it possible to determine computationally whether the drop in RI is only attributed to density change or something else?