Glyoxal Fixation and Its Relationship to Immunohistochemistry

Abstract

Glyoxal is a popular substitute for formalin and in many ways acts like it, although there are significant differences. When formulated correctly, glyoxal fixatives produce superior morphological detail in only 1-9 h, but crosslinking does not occur. Glyoxal has a unique reactivity with arginine, producing a cyclic imidazole in place of the highly charged guanidinium group, thus reducing eosinophilia in arginine-rich tissue elements. In the absence of crosslink-induced masking of epitopes, most antibodies work directly on glyoxal-fixed specimens without the need for antigen retrieval. The arginine reaction does cause loss of immunoreactivity in arginine-rich antigens, however. Fortunately, the imidazole is readily removed by a simple antigen retrieval process: pH 8.6 Tris HCl buffer for 10 min at 125°C. The conformational basis for needing antigen retrieval, and how it works on a molecular level is explained for both glyoxal and formalin fixation. (The J Histotechnol 29:65, 2006)

Submitted November 4, 2005; accepted with revisions March 15, 2006.

Keywords:

• antigen retrieval
• fixation
• formalin
• glyoxal
• HIER
• IHC
• immunohistochemistry
• heat-induced epitope retrieval
• Prefer
• glyoxal-specific antigen retrieval