Solid immersion microscopy images cells under cryogenic conditions with 12 nm resolution

Communications Biology
Lin WangMarisa L Martin-Fernandez

Abstract

Super-resolution fluorescence microscopy plays a crucial role in our understanding of cell structure and function by reporting cellular ultrastructure with 20-30 nm resolution. However, this resolution is insufficient to image macro-molecular machinery at work. A path to improve resolution is to image under cryogenic conditions. This substantially increases the brightness of most fluorophores and preserves native ultrastructure much better than chemical fixation. Cryogenic conditions are, however, underutilised because of the lack of compatible high numerical aperture objectives. Here, using a low-cost super-hemispherical solid immersion lens (superSIL) and a basic set-up we achieve 12 nm resolution under cryogenic conditions, to our knowledge the best yet attained in cells using simple set-ups and/or commercial systems. By also allowing multicolour imaging, and by paving the way to total-internal-reflection fluorescence imaging of mammalian cells under cryogenic conditions, superSIL microscopy opens a straightforward route to achieve unmatched resolution on bacterial and mammalian cell samples.

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Citations

May 2, 2019·Nature Reviews. Microbiology·Ian M Dobbie
Oct 29, 2020·Journal of Microscopy·M L Martin-Fernandez
Sep 2, 2019·Trends in Microbiology·Wen Yang, Ariane Briegel
Nov 20, 2019·Bio-protocol·Benji C BatemanMarisa L Martin-Fernandez
Feb 22, 2021·Journal of Structural Biology·Jie E YangElizabeth R Wright
Jan 15, 2021·Annual Review of Physical Chemistry·Peter D Dahlberg, W E Moerner
Jun 25, 2021·Quarterly Reviews of Biophysics·David J DeRosier
Sep 2, 2021·Optics Letters·Mohammad Hossein KhosraviFaegheh Hajizadeh

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Methods Mentioned

BETA
fluorescence microscopy
electron microscopy
super-resolution microscopy
chip
imaging techniques
optical microscopy

Software Mentioned

STORM
specialist STORM
Elyra
ZEISS ZEN
ZEN
SIL STORM

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