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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., 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.

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Item Permalink: http://hdl.handle.net/11858/00-001M-0000-002E-37C7-C Version Permalink: http://hdl.handle.net/21.11116/0000-0003-B050-B
Genre: Journal Article

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 Creators:
Richter, K. N., Author
Revelo, N. H., Author
Seitz, K. J., Author
Helm, M. S., Author
Sarkar, D., Author
Saleeb, R. S., Author
D'Este, E.1, Author              
Eberle, J., Author
Wagner, E., Author
Vogl, C., Author
Lazaro, D. F., Author
Richter, F., Author
Coy‐Vergara, J., Author
Coceano, G., Author
Boyden, E. S., Author
Duncan, R. R., Author
Hell, S. W.1, Author              
Lauterbach, M. A., Author
Lehnart, S. E., Author
Moser, T.2, Author              
Outeiro, T., AuthorRehling, P.3, Author              Schwappach, B.4, Author              Testa, I., AuthorZapiec, B., AuthorRizzoli, S. O., Author more..
Affiliations:
1Department of NanoBiophotonics, MPI for Biophysical Chemistry, Max Planck Society, ou_578627              
2Research Group of Synaptic Nanophysiology, MPI for Biophysical Chemistry, Max Planck Society, ou_2205655              
3Max Planck Fellow Peter Rehling, ou_1298545              
4Max Planck Fellow Blanche Schwappach, ou_1548137              

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Free keywords: fixation; glyoxal; immunocytochemistry; PFA; super-resolution microscopy
 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.

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Language(s): eng - English
 Dates: 2017-11-162018-01-04
 Publication Status: Published in print
 Pages: -
 Publishing info: -
 Table of Contents: -
 Rev. Method: Peer
 Identifiers: DOI: 10.15252/embj.201695709
 Degree: -

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Title: EMBO Journal
Source Genre: Journal
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Publ. Info: -
Pages: - Volume / Issue: 37 (1) Sequence Number: - Start / End Page: 139 - 159 Identifier: -