Cryo-SOFI enabling low-dose super-resolution correlative light and electron cryo-microscopy.

Proceedings of the National Academy of Sciences of the United States of America
Felipe MoserRainer Kaufmann

Abstract

Correlative light and electron cryo-microscopy (cryo-CLEM) combines information from the specific labeling of fluorescence cryo-microscopy (cryo-FM) with the high resolution in environmental context of electron cryo-microscopy (cryo-EM). Exploiting super-resolution methods for cryo-FM is advantageous, as it enables the identification of rare events within the environmental background of cryo-EM at a sensitivity and resolution beyond that of conventional methods. However, due to the need for relatively high laser intensities, current super-resolution cryo-CLEM methods require cryo-protectants or support films which can severely reduce image quality in cryo-EM and are not compatible with many samples, such as mammalian cells. Here, we introduce cryogenic super-resolution optical fluctuation imaging (cryo-SOFI), a low-dose super-resolution imaging scheme based on the SOFI principle. As cryo-SOFI does not require special sample preparation, it is fully compatible with conventional cryo-EM specimens, and importantly, it does not affect the quality of cryo-EM imaging. By applying cryo-SOFI to a variety of biological application examples, we demonstrate resolutions up to ∼135 nm, an improvement of up to three times compared with conve...Continue Reading

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Citations

Jul 2, 2019·ELife·Sergey GorelickAlex de Marco
Jul 3, 2019·Genes·Udo J Birk
Jul 8, 2020·Annual Review of Virology·Emmanuelle R J QueminKay Grünewald
Feb 23, 2020·Nature Chemical Biology·Daniel Luque, José R Castón
Oct 7, 2020·FEBS Letters·Martin Turk, Wolfgang Baumeister
Nov 4, 2020·Molecular Microbiology·Trishant R UmrekarMorgan Beeby
Nov 23, 2020·Journal of Neurochemistry·Miguel Ricardo Leung, Tzviya Zeev-Ben-Mordehai
Feb 21, 2021·Communications Biology·Sina MangerAchilleas S Frangakis
Feb 22, 2021·Journal of Structural Biology·Jie E YangElizabeth R Wright
Mar 25, 2021·Emerging Topics in Life Sciences·Anshul AssaiyaJanesh Kumar
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Aug 14, 2021·Journal of Molecular Biology·Felix J B Bäuerlein, Wolfgang Baumeister
Sep 8, 2021·Accounts of Chemical Research·Michael ElbaumPeter Rez
Jan 29, 2022·Advanced Science·Jiwoong KwonSang-Hee Shim

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Methods Mentioned

BETA
imaging techniques
FCS
Transfection

Software Mentioned

Unblur
IMOD Etomo
MATLAB
DIPimage toolbox
Bsoft
SOFI

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