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

MPG-Autoren
http://pubman.mpdl.mpg.de/cone/persons/resource/persons98972

D'Este,  E.
Department of NanoBiophotonics, MPI for Biophysical Chemistry, Max Planck Society;

http://pubman.mpdl.mpg.de/cone/persons/resource/persons15210

Hell,  S. W.
Department of NanoBiophotonics, MPI for Biophysical Chemistry, Max Planck Society;

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Volltexte (frei zugänglich)

2504420.pdf
(Verlagsversion), 5MB

Ergänzendes Material (frei zugänglich)

2504420_Suppl_1.pdf
(Ergänzendes Material), 13MB

2504420_Suppl_3.pdf
(Ergänzendes Material), 3MB

Zitation

Richter, K. N., Revelo, N. H., Seitz, K. J., Helm, M. S., Sarkar, D., Saleeb, R. S., et al. (2018). Glyoxal as an alternative fixative to formaldehyde in immunostaining and super-resolution microscopy. EMBO Journal, 37(1), 139-159. doi:10.15252/embj.201695709.


Zitierlink: http://hdl.handle.net/11858/00-001M-0000-002E-37C7-C
Zusammenfassung
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.