All-optical Super-resolution Imaging of Molecules in Their Nanoscale Cellular Context

Session

Plenary Talk: Joerg Bewersdorf - All-optical Super-resolution Imaging of Molecules in Their Nanoscale Cellular Context

Authors

Dr. Joerg Bewersdorf (1)

Affiliations

1. Yale University

Keywords

super-resolution, DNA-PAINT, Expansion Microscopy, 4Pi

Abstract text

Super-resolution optical microscopy has become a powerful tool to study the nanoscale spatial distribution of molecules of interest in biological cells, tissues and other structures over the last years. Imaging these distributions in the context of other molecules or the general structural context is, however, still challenging. I will present recent developments of our lab focusing on instrumentation, fluorescent probes and sample preparation techniques which tackle this challenge. 4Pi-SMS microscopy, an interferometrical single-molecule localization technique, simultaneously localizes up to three species of proteins with sub-10 nm localization precision in 3D [1]. 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 [2]. Labeling proteins and other cellular components in bulk in our recent pan-Expansion Microscopy method provides ultrastructural context to the nanoscale organization of proteins, replacing complex correlative light/electron microscopy by an all-optical imaging approach [3,4,5].  

Financial Interest Disclosure: J.B. has financial interest in Bruker Corp. and Hamamatsu Photonics and is co-founder of panluminate, a startup company related to Expansion Microscopy. 

References

[1] Zhang, Y., Schroeder, L.K., et al. “Fluorogenic probe for fast 3D whole-cell DNA-PAINT”. Nature Methods (2020). https://doi.org/10.1038/s41592-019-0676-4  

[2] Chung, K.K.H. et al. “Fluorogenic DNA-PAINT for faster, low-background super-resolution imaging”. Nature Methods (2022). https://doi.org/10.1038/s41592-022-01464-9
 [3] M’Saad, O., Bewersdorf, J. “Light microscopy of proteins in their ultrastructural context”. Nature Communications (2020). https://doi.org/10.1038/s41467-020-17523-8

[4] M’Saad, O., et al. “All-optical visualization of specific molecules in the ultrastructural context of brain tissue”. bioRxiv (2022). https://doi.org/10.1101/2022.04.04.486901 

[4] M’Saad, O., et al. “Unclearing Microscopy”. bioRxiv (2022). doi: https://doi.org/10.1101/2022.11.29.518361