Glyoxal as an alternative fixative to formaldehyde in immunostaining and super-resolution microscopy
2018 | journal article; research paper
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Glyoxal as an alternative fixative to formaldehyde in immunostaining and super-resolution microscopy
Richter, K. N. ; Revelo, N. H. ; Seitz, K. J.; Helm, M. S.; Sarkar, D.; Saleeb, R. S. & d’Este, E. et al. (2018)
The EMBO journal, 37(1) pp. 139-159. DOI: https://doi.org/10.15252/embj.201695709
Documents & Media
Details
- 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