Multiview confocal super-resolution microscopy

Autor: Yicong Wu, Ryan Christensen, Arpita Upadhyaya, Jiamin Liu, Yilun Sun, Akshay Patel, Titas Sengupta, Yves Pommier, Hari Shroff, Ivan Rey-Suarez, Jonathan S. Daniels, Christian A. Combs, Daniel A. Colón-Ramos, Lingyu Bao, Jiji Chen, Melissa Glidewell, Junhui Sun, Xufeng Wu, Robert S. Fischer, Xiaofei Han, Corey Smith, Leighton H. Duncan, Yun-Bo Shi, Sougata Roy, Yijun Su, Elizabeth Murphy, Patrick J. La Riviere
Rok vydání: 2021
Předmět:
Zdroj: Nature. 600:279-284
ISSN: 1476-4687
0028-0836
Popis: Confocal microscopy1 remains a major workhorse in biomedical optical microscopy owing to its reliability and flexibility in imaging various samples, but suffers from substantial point spread function anisotropy, diffraction-limited resolution, depth-dependent degradation in scattering samples and volumetric bleaching2. Here we address these problems, enhancing confocal microscopy performance from the sub-micrometre to millimetre spatial scale and the millisecond to hour temporal scale, improving both lateral and axial resolution more than twofold while simultaneously reducing phototoxicity. We achieve these gains using an integrated, four-pronged approach: (1) developing compact line scanners that enable sensitive, rapid, diffraction-limited imaging over large areas; (2) combining line-scanning with multiview imaging, developing reconstruction algorithms that improve resolution isotropy and recover signal otherwise lost to scattering; (3) adapting techniques from structured illumination microscopy, achieving super-resolution imaging in densely labelled, thick samples; (4) synergizing deep learning with these advances, further improving imaging speed, resolution and duration. We demonstrate these capabilities on more than 20 distinct fixed and live samples, including protein distributions in single cells; nuclei and developing neurons in Caenorhabditis elegans embryos, larvae and adults; myoblasts in imaginal disks of Drosophila wings; and mouse renal, oesophageal, cardiac and brain tissues. A combination of multiview imaging, structured illumination, reconstruction algorithms and deep-learning predictions realizes spatial- and temporal-resolution improvements in fluorescence microscopy to produce super-resolution images from diffraction-limited input images.
Databáze: OpenAIRE
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