BIOTECHNOLOGY 57
Staining specimens allows researchers to cast a broader net for investigating targets.
The advantages of staining specimens
A look at developments in multicolour enzymatic immunohistochemistry assays for formalin-fixed paraffin-embedded tissue.
Regard sur les développements liés aux analyses immunohistochimiques d’enzymes multicolores pour les tissus fixés au formol et incrustés dans la paraffine.
Ein Blick auf die Entwicklungen mehrfarbiger enzymatischer Immunohistochemie-Assays für Formalin-fixiertes Paraffin- eingebettetes Gewebe.
I
mmunofluorescence (IF) has become a preferred method of concurrently detecting multiple antigenic markers within a single tissue specimen. However, immunoenzymatic chromogen staining with multiple coloured substrates remains an informative and important research tool.
Staining specimens with immunoenzymatic chromogens allows researchers to cast a broader net for investigating targets because, unlike IF, it is permanent and can be visualised in relation to the comprehensive morphology of tissue specimens.
Tis stability also allows standard histological stains to be used in conjunction with the immunohistochemistry (IHC) to give researchers an additional layer of information.
Immunofluorescence is often preferred over enzymatic IHC because it is a technically simpler method of visualising multiple antigenic markers.
Imaging with a fluorescent microscope and creating the composite images of multiple IF colour channels can be the most complicated aspect of IF staining, but quantification of
distinctly stained elements is simple and precise.
Te development of assays involving multiple IHC chromogenic substrates presents many challenges, such as determining the appropriate sequence of marker application/ detection, compatibility of cellular localisation of combined markers, special requirements for preparation of various enzymatic substrates, visual contrast compatibility of chromogenic substrates, the length of the overall staining process, and methods of analysing staining results.
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