Immunohistochemical analysis of formaldehyde- and trypsin- or pepsin-treated material

Abstract

Immunohistochemical studies of paraffin sections of formaldehyde-fixed material and formaldehyde-fixed cryostat sections treated with trypsin or other enzymes were undertaken. Formaldehyde treatment resulted mostly in partial or total loss of antigenicity, as visualized by fluorescent antibody technique in tissue sections. The digestion of sections with trypsin resulted usually in total or partial restitution of antigenicity, previously altered with formaldehyde. This effect is suggested to consist primarily in breaking formaldehyde-induced intermolecular cross-links. Restitution of antigenicity altered by formaldehyde can also be achieved on using pepsin or other proteolytic enzymes, or by some other technique. When the effect of one enzyme proves insufficient, it can readily be complemented by the action of another enzyme applied subsequently. Mild trypsin treatment preserves well the morphological structure of most tissues.