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Glyoxal as an alternative fixative to formaldehyde in immunostaining and super‐resolution microscopy
Name
embj.201695709.pdf
Description
Published version
Size
4.81 MB
Format
Adobe PDF
Checksum (MD5)
8254169afac961fc6f2bc38d15a32035
Author(s)
Richter, Katharina N
Revelo, Natalia H
Seitz, Katharina J
Helm, Martin S
Sarkar, Deblina
Saleeb, Rebecca S
D'Este, Elisa
Eberle, Jessica
Wagner, Eva
Vogl, Christian
Date Issued
2018
Journal
EMBO Journal, The
Publisher
EMBO
Version
Final published version
Abstract
© 2017 The Authors. Published under the terms of the CC BY 4.0 license Paraformaldehyde (PFA) is the most commonly used fixative for immunostaining of cells, but has been associated with various problems, ranging from loss of antigenicity to changes in morphology during fixation. We show here that the small dialdehyde glyoxal can successfully replace PFA. Despite being less toxic than PFA, and, as most aldehydes, likely usable as a fixative, glyoxal has not yet been systematically tried in modern fluorescence microscopy. Here, we tested and optimized glyoxal fixation and surprisingly found it to be more efficient than PFA-based protocols. Glyoxal acted faster than PFA, cross-linked proteins more effectively, and improved the preservation of cellular morphology. We validated glyoxal fixation in multiple laboratories against different PFA-based protocols and confirmed that it enabled better immunostainings for a majority of the targets. Our data therefore support that glyoxal can be a valuable alternative to PFA for immunostaining.
Terms of Use
Creative Commons Attribution 4.0 International license
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DOI of Published Version
10.15252/EMBJ.201695709
