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  • 1. Richter, Katharina N.
    et al.
    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
    KTH, Centres, Science for Life Laboratory, SciLifeLab. KTH, School of Engineering Sciences (SCI), Applied Physics.
    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
    KTH, School of Engineering Sciences (SCI), Applied Physics. KTH, Centres, Science for Life Laboratory, SciLifeLab.
    Zapiec, Bolek
    Rizzoli, Silvio O.
    Glyoxal as an alternative fixative to formaldehyde in immunostaining and super-resolution microscopy2018In: EMBO Journal, ISSN 0261-4189, E-ISSN 1460-2075, Vol. 37, no 1, p. 139-159Article in journal (Refereed)
    Abstract [en]

    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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