title: A simple and reliable immunohistochemical method for colocalization of 2 antigens in the same cells of paraffin-embedded tissues.
abstract: Immunohistochemistry (IHC) lacks an efficient technique for colocalizing multiple antigens in the same cells of a single tissue section. The development of a methodology which combines the advantage of low cost, high sensitivity, and specificity would benefit clinical diagnosis and general research. On the basis of a newly published method of visualizing 2 antigens on a single paraffin-embedded tissue section, we have further developed a novel sequential technique for colocalizing 2 different antigens in a same cell in a paraffin-embedded tissue section. In this technique, we combined the microwave heating technique (MVT) with normal IHC methods to sequentially double stain a paraffin section; and colocalize 2 antigens in a single cell through result comparison stored in a digital management system. This MVT colocalization method has a higher degree of sensitivity and specificity comparable with conventional staining of both immunofluorescence and IHC systems. The primary advantage of this method is that it is inexpensive and convenient; the antibody(s) used in this method can be generated from the same or different species; it allows colocalization or comparison of different results of cell morphology for any single cell of the section on 2 images, avoids uncertainty when overlapping 2 antigens on a single image.
authors.affiliation: UQ Centre for Clinical Research, University of Queensland, Brisbane, Qld, Australia.
authors.forename: Jun
authors.initials: J
authors.lastname: Yan
authors.forename: Vibeke S
authors.initials: VS
authors.lastname: Catts
authors.forename: Anthony
authors.initials: A
authors.lastname: Chan
authors.forename: Pamela A
authors.initials: PA
authors.lastname: McCombe
mesh_terms: D000941:Antigens / Q000276:immunology; D006361:Heating; D006801:Humans; D007150:Immunohistochemistry / Q000295:instrumentation / Q000379:methods*; D008872:Microwaves; D016612:Paraffin Embedding
chemical_list: D000941:Antigens