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Journal Article

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

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

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

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Moser,  T.
Research Group of Synaptic Nanophysiology, MPI for Biophysical Chemistry, Max Planck Society;

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Fulltext (public)

2504420.pdf
(Publisher version), 5MB

Supplementary Material (public)

2504420_Suppl_1.pdf
(Supplementary material), 13MB

2504420_Suppl_3.pdf
(Supplementary material), 3MB

Citation

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.


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