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

2018 | journal article; research paper

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​Richter, K. N., Revelo, N. H., Seitz, K. J., Helm, M. S., Sarkar, D., Saleeb, R. S., d’Este, E. ... Rizzoli, S. O. (2018). ​Glyoxal as an alternative fixative to formaldehyde in immunostaining and super-resolution microscopy. The EMBO journal37(1), ​139​-159​. ​doi: https://doi.org/10.15252/embj.201695709 

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Authors
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 ; Lazaro, Diana F. ; Richter, Frank ; Coy-Vergara, 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. 
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.
Issue Date
2018
Journal
The EMBO journal 
Project
SFB 1002: Modulatorische Einheiten bei Herzinsuffizienz 
SFB 1002 | A09: Lokale molekulare Nanodomänen-Regulation der kardialen Ryanodin-Rezeptor-Funktion 
SFB 1190: Transportmaschinen und Kontaktstellen zellulärer Kompartimente 
SFB 1190 | P09: Proteinsortierung in der Synapse: Prinzipien und molekulare Organisation 
Working Group
RG Hell 
RG Lehnart (Cellular Biophysics and Translational Cardiology Section) 
RG Rehling (Mitochondrial Protein Biogenesis) 
RG Schwappach (Membrane Protein Biogenesis) 
RG Rizzoli (Quantitative Synaptology in Space and Time) 
External URL
https://sfb1002.med.uni-goettingen.de/production/literature/publications/195
https://sfb1190.med.uni-goettingen.de/production/literature/publications/15
eISSN
1460-2075
Language
English

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