Journal
EMBO JOURNAL
Volume 37, Issue 1, Pages 139-159Publisher
WILEY
DOI: 10.15252/embj.201695709
Keywords
fixation; glyoxal; immunocytochemistry; PFA; super-resolution Microscopy
Categories
Funding
- Deutsche Forschungsgemeinschaft through the Collaborative Research Center [889, 1002, 1190]
- European Research Council [ERC-CoG-614765]
- ERC [ERCAdG 339580, 638314]
- Boehringer Ingelheim Fonds
- ITN TAMPting network (People Programme (Marie Curie Actions) of the European Union's Seventh Framework Programme (FP7) under the Research Executive Agency) [607072]
- EMBO Long-Term Fellowship [ALTF 232-2016]
- Netherlands Organization for Scientific Research [NWO-ALW 016.Veni.171.097]
- German Center for Cardiovascular Research, DZHK
- Open Philanthropy project
- HHMI-Simons Faculty Scholars Program
- US-Israel Binational Science Foundation [2014509]
- Medical Research Council [MR/K01563X/1] Funding Source: researchfish
- European Research Council (ERC) [638314] Funding Source: European Research Council (ERC)
- MRC [MR/K01563X/1] Funding Source: UKRI
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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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