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

• Katharina N Richter ^[1] ; Natalia H Revelo ^[1] ; Katharina J Seitz ^[3] ; Martin S Helm ^[3] ; Deblina Sarkar ^[4] ; Rebecca S Saleeb ^[5] ; Elisa D'Este ^[6] ; Jessica Eberle ^[7] ; Eva Wagner ^[8] ; Christian Vogl ^[9] ; Diana F Lazaro ^[10] ; Frank Richter ^[11] ; Javier Coy‐Vergara ^[12] ; Giovanna Coceano ^[2] ; Edward S Boyden ^[13] ; Rory R Duncan ^[5] ; Stefan W Hell ^[6] ; Marcel A Lauterbach ^[7] ; Stephan E Lehnart ^[8] ; Tobias Moser ^[9] ; Tiago F Outeiro ^[10] ; Peter Rehling ^[14] ; Blanche Schwappach ^[12] ; Ilaria Testa ^[2] ; Bolek Zapiec ^[15] ; Silvio O Rizzoli ^[1]
  1. [1] University of Göttingen

     University of Göttingen

     Landkreis Göttingen, Alemania

     
  2. [2] Royal Institute of Technology

     Royal Institute of Technology

     Suecia

     
  3. [3] 1 Department of Neuro‐ and Sensory Physiology University of Göttingen Medical Center Göttingen Germany; 3 International Max Planck Research School Molecular Biology Göttingen Germany
  4. [4] 4 MIT Media Lab
  5. [5] 5 Edinburgh Super‐Resolution Imaging Consortium Institute of Biological Chemistry, Biophysics, and Bioengineering Heriot‐Watt University Edinburgh UK
  6. [6] 6 Department of NanoBiophotonics Max‐Planck‐Institute for Biophysical Chemistry Göttingen Germany
  7. [7] 7 Department of Neural Systems Max‐Planck‐Institute for Brain Research Frankfurt am Main Germany
  8. [8] 8 Heart Research Center Göttingen Department of Cardiology & Pulmonology University Medical Center Göttingen Göttingen Germany; 9 German Center for Cardiovascular Research (DZHK) Site Göttingen
  9. [9] 10 Institute for Auditory Neuroscience and InnerEarLab University Medical Center Göttingen Göttingen Germany; 11 Max‐Planck‐Institute for Experimental Medicine Auditory Neuroscience Group Göttingen Germany
  10. [10] 12 Department of Experimental Neurodegeneration Center for Nanoscale Microscopy and Molecular Physiology of the Brain Center for Biostructural Imaging of Neurodegeneration University Medical Center Göttingen Göttingen Germany; 13 Max‐Planck‐Institute for Experimental Medicine Göttingen Germany
  11. [11] 3 International Max Planck Research School Molecular Biology Göttingen Germany; 14 Department of Cellular Biochemistry University Medical Center Göttingen Göttingen Germany
  12. [12] 15 Department of Molecular Biology University Medical Center Göttingen Göttingen Germany
  13. [13] 17 Departments of Brain and Cognitive Science and Biological Engineering MIT Media Lab and McGovern Institute Cambridge MA USA
  14. [14] 14 Department of Cellular Biochemistry University Medical Center Göttingen Göttingen Germany; 18 Max‐Planck‐Institute for Biophysical Chemistry Göttingen Germany
  15. [15] 19 Max Planck Research Unit for Neurogenetics Frankfurt am Main Germany

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• Localización: EMBO journal: European Molecular Biology Organization, ISSN 0261-4189, Vol. 37, Nº. 1, 2018, págs. 139-159
• Idioma: inglés
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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.