Could Glyoxal fixation enhance immunohistochemical signalling?
May 16，2024
Yes. Glyoxal fixation enhances immunohistochemical signals, and adjusting the glyoxal fixative to pH 4 or 5 is key for this effect. Glyoxal fixation typically enhances immune signals compared to formaldehyde and glutaraldehyde. This may be due to the fact that the chemical structure of glyoxal gives it good cross-linking properties that preserve the structure and antigenicity of proteins, making it suitable for immunohistochemistry. Studies have also shown that ethanol-free glyoxal fixatives enhanced many synaptic molecules more than glyoxal fixatives containing 20% ethanol. Increasing the concentrations of glyoxal and acetic acid to 9% and 8%, respectively, also resolves the problem of tissue section fragility.
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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.
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