Advances in immunocytochemistry.
Recent advances in immunohisto/cytochemical methods have been directed towards new fluorescent labels, and to increasing sensitivity and improving methods for multiple immunostaining. The newest fluorescent dyes come in many colours, are much more stable than fluorescein isothiocyanate and give brighter fluorescence. The greatest impact on sensitivity comes from heat-induced antigen retrieval on paraffin sections. Biotinylated tyramide can also increase enormously the amount of label on a preparation. The immunogold method with silver enhancement has been improved by nanogold, which is smaller than colloidal gold, attracts more silver and increases the sensitivity even further. The difficulties of multiple labelling with two or more primary antibodies raised in the same species due to cross-binding between reagents have largely been overcome. There are new methods of blocking spare reactive sites on the first reaction by strong heat, e.g. from microwaving in buffer. Alternatively, one of the antibodies can be directly biotinylated very easily with a new commercially available kit.
Van Noorden
Susan
S
Department of Histopathology, Imperial College Faculty of Medicine, Hammersmith Hospital, London, UK. s.vannoorden@ic.ac.uk
D000818:Animals; D000906:Antibodies; D000918:Antibody Specificity; D004396:Coloring Agents; D006801:Humans; D007150:Immunohistochemistry / Q000379:methods / Q000639:trends*; D007202:Indicators and Reagents; D016610:Tissue Embedding
D000906:Antibodies; D004396:Coloring Agents; D007202:Indicators and Reagents