Glyoxal Fixation as an Alternative for Zebrafish Embryo Immunostaining

Methods Mol Biol. 2021:2218:245-252. doi: 10.1007/978-1-0716-0970-5_19.


Immunohistochemistry has been widely used as a robust technique to determine the cellular and subcellular localization of proteins. This information ultimately helps to understand the function of these proteins and how biological processes are regulated. Antibodies applicable for labeling in zebrafish are limited, making immuno-staining challenging. Recently glyoxal fixation was rediscovered in tissue culture, mouse, rat, and Drosophila, expanding the list of effective antibodies for these species. Here, we compare a protocol for zebrafish staining using glyoxal as a fixative agent with PFA. We demonstrate that glyoxal fixation improves the antigenicity of some epitopes thereby increasing the number of useful antibodies in zebrafish.

Keywords: Bucky ball; Fixation method; Germ plasm; Glyoxal; Immunostaining; PFA; Zebrafish.

MeSH terms

  • Animals
  • Antibodies / metabolism
  • Embryo, Mammalian / metabolism*
  • Female
  • Fixatives / metabolism
  • Glyoxal / metabolism*
  • Immunohistochemistry / methods*
  • Male
  • Staining and Labeling / methods
  • Tissue Fixation / methods*
  • Zebrafish / metabolism*
  • Zebrafish Proteins / metabolism


  • Antibodies
  • Fixatives
  • Zebrafish Proteins
  • Glyoxal