Toggle Main Menu Toggle Search

Open Access padlockePrints

Glyoxal as an alternative fixative to formaldehyde in immunostaining and super-resolution microscopy

Lookup NU author(s): Professor Tiago OuteiroORCiD

Downloads


Licence

This work is licensed under a Creative Commons Attribution 4.0 International License (CC BY 4.0).


Abstract

© 2017 EMBO.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.


Publication metadata

Author(s): Richter KN, Revelo NH, Seitz KJ, Helm MS, Sarkar D, Saleeb RS, D'Este E, Eberle J, Wagner E, Vogl C, Lazaro DF, Richter F, Coy-Vergara J, Coceano G, Boyden ES, Duncan RR, Hell SW, Lauterbach MA, Lehnart SE, Moser T, Outeiro T, Rehling P, Schwappach B, Testa I, Zapiec B, Rizzoli SO

Publication type: Article

Publication status: Published

Journal: EMBO Journal

Year: 2017

Online publication date: 16/11/2017

Acceptance date: 06/10/2017

Date deposited: 20/12/2017

ISSN (print): 0261-4189

ISSN (electronic): 1460-2075

Publisher: Wiley-VCH Verlag

URL: https://doi.org/10.15252/embj.201695709

DOI: 10.15252/embj.201695709


Altmetrics

Altmetrics provided by Altmetric


Funding

Funder referenceFunder name
607072
ERCAdG 339580
ERC-CoG-614765

Share