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

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 TF, Rehling P, Schwappach B, Testa I, Zapiec B, Rizzoli SO

EMBO J. 37 (1) 139-159 [2018-01-04; online 2017-11-16]

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.

Affiliated researcher

Fellows programme

Ilaria Testa

PubMed 29146773

DOI 10.15252/embj.201695709

Crossref 10.15252/embj.201695709

pii: embj.201695709
pmc: PMC5753035


Publications 7.1.2