Imaging enzymes at work: metabolic mapping by enzyme histochemistry.
For the understanding of functions of proteins in biological and pathological processes, reporter molecules such as fluorescent proteins have become indispensable tools for visualizing the location of these proteins in intact animals, tissues, and cells. For enzymes, imaging their activity also provides information on their function or functions, which does not necessarily correlate with their location. Metabolic mapping enables imaging of activity of enzymes. The enzyme under study forms a reaction product that is fluorescent or colored by conversion of either a fluorogenic or chromogenic substrate or a fluorescent substrate with different spectral characteristics. Most chromogenic staining methods were developed in the latter half of the twentieth century but still find new applications in modern cell biology and pathology. Fluorescence methods have rapidly evolved during the last decade. This review critically evaluates the methods that are available at present for metabolic mapping in living animals, unfixed cryostat sections of tissues, and living cells, and refers to protocols of the methods of choice.
Van Noorden
Cornelis J F
CJ
Department of Cell Biology and Histology, Academic Medical Center, Meibergdreef 15, 1105 AZ Amsterdam, The Netherlands. c.j.vannoorden@amc.uva.nl
D000818:Animals; D003585:Cell Biology; D004798:Enzymes / Q000235:genetics / Q000378:metabolism*; D005456:Fluorescent Dyes; D005910:Glioma / Q000201:enzymology / Q000473:pathology; D006651:Histocytochemistry / Q000379:methods*; D006801:Humans; D007700:Kinetics; D008156:Luciferases / Q000235:genetics / Q000378:metabolism; D008853:Microscopy / Q000379:methods; D010336:Pathology / Q000379:methods / Q000639:trends; D010455:Peptides / Q000378:metabolism; D012333:RNA, Messenger / Q000235:genetics; D013053:Spectrophotometry / Q000379:methods
D004798:Enzymes; D005456:Fluorescent Dyes; D010455:Peptides; D012333:RNA, Messenger; D008156:Luciferases