All-optical imaging of molecules in their nanoscale cellular context
Pr. Joerg Bewesdorf
Yale University School of Medicine
Super-resolution microscopy has become a powerful tool to study the nanoscale spatial distribution of proteins of interest in cells over the last years. Imaging any of these distributions in the context of other proteins or the general cellular context is, however, still challenging. In this webinar, I will present recent developments of our lab which offer a diverse set of solutions that tackle this challenge: ‘Salvaged fluorescence’ enables ratiometric 3-color single-molecule super-resolution imaging at 5–10-nm localization precision in all color channels at negligible chromatic shift or cross-talk . A new fluorogenic DNA-PAINT probe enables fast, 3D whole-cell imaging without the need for optical sectioning, adding a versatile tool to the toolbox of single-molecule super-resolution probes . Labeling proteins and other cellular components in bulk in our novel pan-Expansion Microscopy method provides ultrastructural context to the nanoscale organization of proteins, replacing complex correlative light/electron microscopy with an all-optical imaging approach .
 Zhang, Y., Schroeder, L.K. et al. “Nanoscale subcellular architecture revealed by multicolor three-dimensional salvaged fluorescence imaging”. Nat Methods 17, 225–231 (2020).
 Chung, K.K.H. et al. “Fluorogenic probe for fast 3D whole-cell DNA-PAINT”. bioRxiv (2020).
 M’Saad, O. and Bewersdorf, J. “Light microscopy of proteins in their ultrastructural context”.
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