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

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Glyoxal as an alternative fixative to formaldehyde in immunostaining and super-resolution microscopy. / Richter, Katharina N; 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; 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; Zapiec, Bolek; Rizzoli, Silvio O.

In: EMBO J, Vol. 37, No. 1, 04.01.2018, p. 139-159.

Research output: SCORING: Contribution to journal › SCORING: Journal article › Research › peer-review

Harvard

Richter, KN, Revelo, NH, Seitz, KJ, Helm, MS, Sarkar, D, Saleeb, RS, D'Este, E, Eberle, J, Wagner, E, Vogl, C, Lazaro, DF, Richter, F, Coy-Vergara, J, Coceano, G, Boyden, ES, Duncan, RR, Hell, SW, Lauterbach, MA, Lehnart, SE, Moser, T, Outeiro, TF, Rehling, P, Schwappach, B, Testa, I, Zapiec, B & Rizzoli, SO 2018, 'Glyoxal as an alternative fixative to formaldehyde in immunostaining and super-resolution microscopy', EMBO J, vol. 37, no. 1, pp. 139-159. https://doi.org/10.15252/embj.201695709

APA

Richter, K. N., Revelo, N. H., Seitz, K. J., Helm, M. S., Sarkar, D., Saleeb, R. S., D'Este, E., Eberle, J., Wagner, E., Vogl, C., Lazaro, D. F., Richter, F., Coy-Vergara, J., Coceano, G., Boyden, E. S., Duncan, R. R., Hell, S. W., Lauterbach, M. A., Lehnart, S. E., ... Rizzoli, S. O. (2018). Glyoxal as an alternative fixative to formaldehyde in immunostaining and super-resolution microscopy. EMBO J, 37(1), 139-159. https://doi.org/10.15252/embj.201695709

Vancouver

Richter KN, Revelo NH, Seitz KJ, Helm MS, Sarkar D, Saleeb RS et al. Glyoxal as an alternative fixative to formaldehyde in immunostaining and super-resolution microscopy. EMBO J. 2018 Jan 4;37(1):139-159. https://doi.org/10.15252/embj.201695709

Bibtex

@article{834858c42165491984f24dba366a749b,
title = "Glyoxal as an alternative fixative to formaldehyde in immunostaining and 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.",
keywords = "Animals, COS Cells, Chlorocebus aethiops, Drosophila melanogaster, Fixatives/chemistry, Formaldehyde/chemistry, Glyoxal/chemistry, HeLa Cells, Humans, Immunohistochemistry/methods, Mice, Microscopy, Fluorescence/methods, Nerve Tissue Proteins/metabolism, Tissue Fixation/methods",
author = "Richter, {Katharina N} and Revelo, {Natalia H} and Seitz, {Katharina J} and Helm, {Martin S} and Deblina Sarkar and Saleeb, {Rebecca S} and Elisa D'Este and Jessica Eberle and Eva Wagner and Christian Vogl and Lazaro, {Diana F} and Frank Richter and Javier Coy-Vergara and Giovanna Coceano and Boyden, {Edward S} and Duncan, {Rory R} and Hell, {Stefan W} and Lauterbach, {Marcel A} and Lehnart, {Stephan E} and Tobias Moser and Outeiro, {Tiago F} and Peter Rehling and Blanche Schwappach and Ilaria Testa and Bolek Zapiec and Rizzoli, {Silvio O}",
note = "{\textcopyright} 2017 The Authors. Published under the terms of the CC BY 4.0 license.",
year = "2018",
month = jan,
day = "4",
doi = "10.15252/embj.201695709",
language = "English",
volume = "37",
pages = "139--159",
journal = "EMBO J",
issn = "0261-4189",
publisher = "NATURE PUBLISHING GROUP",
number = "1",

}

RIS

TY - JOUR

T1 - Glyoxal as an alternative fixative to formaldehyde in immunostaining and super-resolution microscopy

AU - Richter, Katharina N

AU - Revelo, Natalia H

AU - Seitz, Katharina J

AU - Helm, Martin S

AU - Sarkar, Deblina

AU - Saleeb, Rebecca S

AU - D'Este, Elisa

AU - Eberle, Jessica

AU - Wagner, Eva

AU - Vogl, Christian

AU - Lazaro, Diana F

AU - Richter, Frank

AU - Coy-Vergara, Javier

AU - Coceano, Giovanna

AU - Boyden, Edward S

AU - Duncan, Rory R

AU - Hell, Stefan W

AU - Lauterbach, Marcel A

AU - Lehnart, Stephan E

AU - Moser, Tobias

AU - Outeiro, Tiago F

AU - Rehling, Peter

AU - Schwappach, Blanche

AU - Testa, Ilaria

AU - Zapiec, Bolek

AU - Rizzoli, Silvio O

N1 - © 2017 The Authors. Published under the terms of the CC BY 4.0 license.

PY - 2018/1/4

Y1 - 2018/1/4

N2 - 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.

AB - 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.

KW - Animals

KW - COS Cells

KW - Chlorocebus aethiops

KW - Drosophila melanogaster

KW - Fixatives/chemistry

KW - Formaldehyde/chemistry

KW - Glyoxal/chemistry

KW - HeLa Cells

KW - Humans

KW - Immunohistochemistry/methods

KW - Mice

KW - Microscopy, Fluorescence/methods

KW - Nerve Tissue Proteins/metabolism

KW - Tissue Fixation/methods

U2 - 10.15252/embj.201695709

DO - 10.15252/embj.201695709

M3 - SCORING: Journal article

C2 - 29146773

VL - 37

SP - 139

EP - 159

JO - EMBO J

JF - EMBO J

SN - 0261-4189

IS - 1

ER -