Collagenase predigestion on paraffin sections enhances collagen immunohistochemical detection without distorting tissue morphology.
Reliable immunohistochemical detection of collagen in formalin fixed, paraffin embedded tissues requires protease digestion. While these pan-proteases (pepsin, trypsin, protease K, etc.) enhance collagen detection, they also digest many other tissue proteins and produce poor cellular morphology and unrecognizable cellular structures. Balancing the conditions (protease type, concentration, incubation time and temperature) to digest some, but not all, proteins in a tissue section while optimizing collagen detection requires one to compromise improved collagen immunolabeling with adequate cellular morphology. Furthermore, optimal conditions for digesting tissue proteins to enhance collagen detection vary among tissue types and their fixation. Although brain is not typically subject to these deleterious consequences, structures such as epithelium, spermatids, stroma etc. and other tissues with complicated histology are profoundly affected. To resolve this technical dilemma, we discovered a novel use for collagenase to enhance collagen immunodetection without affecting the noncollagen proteins, thereby preserving tissue morphology. Collagenase, which is typically used in vitro for disassociation of cells, has never been used reliably on formalin fixed, paraffin embedded tissue sections. This new use of collagenase for immunohistochemistry promotes increased collagen immunolabeling, is easy to use, is versatile, and allows preservation of tissue structure that provides maximal and accurate histological information.
D'Andrea
M R
MR
Johnson and Johnson Pharmaceutical Research and Development, Drug, Discovery, Spring House, PA 19477, USA. mdandrea@prdus.jnj.com
D048429:Cell Size; D002478:Cells, Cultured / Q000166:cytology* / Q000378:metabolism*; D003094:Collagen / Q000378:metabolism* / Q000648:ultrastructure*; D017364:Collagenases / Q000378:metabolism*; D006801:Humans; D007150:Immunohistochemistry; D016612:Paraffin Embedding / Q000379:methods*; D013194:Staining and Labeling / Q000379:methods*
D003094:Collagen; D017364:Collagenases