Single-shot label-free nanoscopy for quantitative organelle visualization on standard commercial microscopes

Inutsuka, Y.; Yamamoto, K.; Kuroda, M.; Okada, Y.

doi:10.1101/2024.12.31.630894

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bioRxiv · 10.1101/2024.12.31.630894

Single-shot label-free nanoscopy for quantitative organelle visualization on standard commercial microscopes

Inutsuka, Y.·Yamamoto, K.·Kuroda, M.·Okada, Y.

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Abstract

Label-free microscopy techniques that overcome the limitations of fluorescence microscopy are increasingly needed in biological research. Quantitative phase imaging has emerged as a powerful label-free method for live cell imaging, particularly at multicellular scales. However, a critical gap exists in achieving easy-to-implement, high-speed, high-resolution microscopy at subcellular scales. Here, we report upgraded polarization-resolved differential phase contrast nanoscopy (UpDPC); this single-shot computational phase microscopy (CPM) enhances resolution and sensitivity to subcellular levels. UpDPC can be readily integrated into commercial fluorescence microscopes, enabling 100 nm gap separation, over 100 Hz imaging via single-shot phase retrieval, and 100 ms computation of phase image during observation. Our results demonstrate that high-performance CPM provides reproducible and comprehensive visualization of various organelles, offering a practical and scalable implementation for basic research and clinical applications.

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Inutsuka, Y., Yamamoto, K., Kuroda, M., Okada, Y.. 2025-01-02. Single-shot label-free nanoscopy for quantitative organelle visualization on standard commercial microscopes. https://doi.org/10.1101/2024.12.31.630894

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