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Lightsheet localization microscopy enables fast, large-scale, and three-dimensional super-resolution imaging
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Lightsheet localization microscopy enables fast, large-scale, and three-dimensional super-resolution imaging

Chieh-Han Lu, Wei-Chun Tang, Yen-Ting Liu, Shu-Wei Chang, Frances Camille M. Wu, Chin-Yi Chen, Yun-Chi Tsai, Shun-Min Yang, Chiung-Wen Kuo, Yasushi Okada, …
Communications Biology, 卷.2(1), 177
12/2019
PMID: 31098410

摘要

Medicine (miscellaneous) Biochemistry Genetics and Molecular Biology (all) Agricultural and Biological Sciences (all)
Recent advances in super-resolution microscopy allow the localization of single molecules within individual cells but not within multiple whole cells due to weak signals from single molecules and slow acquisition process for point accumulation to reconstruct super-resolution images. Here, we report a fast, large-scale, and three-dimensional super-resolution fluorescence microscope based on single-wavelength Bessel lightsheet to selectively illuminate spontaneous blinking fluorophores tagged to the proteins of interest in space. Critical parameters such as labeling density, excitation power, and exposure time were systematically optimized resulting in a maximum imaging speed of 2.7 × 10 4 µm 3 s −1 . Fourier ring correlation analysis revealed a reconstructed image with a lateral resolution of ~75 nm through the accumulation of 250 image volumes on immobilized samples within 15 min. Hence, the designed system could open new insights into the discovery of complex biological structures and live 3D localization imaging.

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https://doi.org/10.1038/s42003-019-0403-9檢視
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