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Glyoxal as an alternative fixative to formaldehyde in immunostaining and super‐resolution microscopy
Author(s) -
Richter Kathari,
Revelo Natalia H,
Seitz Katharina J,
Helm Martin S,
Sarkar Deblina,
Saleeb Rebecca S,
D'Este Elisa,
Eberle Jessica,
Wagner Eva,
Vogl Christian,
Lazaro Diana F,
Richter Frank,
CoyVergara Javier,
Coceano Giovanna,
Boyden Edward S,
Duncan Rory R,
Hell Stefan W,
Lauterbach Marcel A,
Lehnart Stephan E,
Moser Tobias,
Outeiro Tiago F,
Rehling Peter,
Schwappach Blanche,
Testa Ilaria,
Zapiec Bolek,
Rizzoli Silvio O
Publication year - 2017
Publication title -
the embo journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 7.484
H-Index - 392
eISSN - 1460-2075
pISSN - 0261-4189
DOI - 10.15252/embj.201695709
Subject(s) - glyoxal , fixative , immunostaining , paraformaldehyde , glutaraldehyde , context (archaeology) , chemistry , fixation (population genetics) , immunoelectron microscopy , pathology , biology , immunohistochemistry , biochemistry , chromatography , medicine , organic chemistry , cytoplasm , paleontology , gene
Abstract 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.