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Scientific article
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Homogeneous multifocal excitation for high-throughput super-resolution imaging

Published inNature methods, vol. 17, no. 7, p. 726-733
Publication date2020-06-22
First online date2020-06-22
Abstract

Super-resolution microscopies have become an established tool in biological research. However, imaging throughput remains a main bottleneck in acquiring large datasets required for quantitative biology. Here we describe multifocal flat illumination for field-independent imaging (mfFIFI). By integrating mfFIFI into an instant structured illumination microscope (iSIM), we extend the field of view (FOV) to >100 × 100 µm2 while maintaining high-speed, multicolor, volumetric imaging at double the diffraction-limited resolution. We further extend the effective FOV by stitching adjacent images for fast live-cell super-resolution imaging of dozens of cells. Finally, we combine our flat-fielded iSIM with ultrastructure expansion microscopy to collect three-dimensional (3D) images of hundreds of centrioles in human cells, or thousands of purified Chlamydomonas reinhardtii centrioles, per hour at an effective resolution of ~35 nm. Classification and particle averaging of these large datasets enables 3D mapping of posttranslational modifications of centriolar microtubules, revealing differences in their coverage and positioning.

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Citation (ISO format)
MAHECIC, Dora et al. Homogeneous multifocal excitation for high-throughput super-resolution imaging. In: Nature methods, 2020, vol. 17, n° 7, p. 726–733. doi: 10.1038/s41592-020-0859-z
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ISSN of the journal1548-7091
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