A new method to localize acid phosphatase using the confocal laser-scanning microscope.
The aim of the study was to work out a technique for the detection of acid phosphatase enzyme activity by confocal laser-scanning microscope using the histochemical acid phosphatase detection method (after Barka and Anderson 1962, modified by Bowen and Lewis 1985) routinely used for light microscopy. The density and the distribution of enzyme reaction product is dependent on the incubation time, as shown by different confocal images or ELISA reader. The inhibition of the enzyme activity with metal ions shows the same profile known from the literature. This staining method seems to be useful to demonstrate subcellular distribution of the enzyme in the lysosomes and in the Golgi apparatus.
Joint Research Organization of the Hungarian Academy of Sciences and Semmelweis University of Medicine, 1st Institute of Pathology and Experimental Cancer Research, Department of Molecular Pathology, Budapest, Hungary. Szezso@korb1.sote.hu
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Szegedi
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Amin
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Szende
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Bowen
D000135:Acid Phosphatase / Q000032:analysis*; D000165:Acridine Orange; D000818:Animals; D002412:Cations / Q000494:pharmacology; D004396:Coloring Agents; D004797:Enzyme-Linked Immunosorbent Assay; D005456:Fluorescent Dyes; D006416:Hematoxylin; D006801:Humans; D006825:Hybridomas; D007834:Lasers; D008739:Methyl Green; D051379:Mice; D018613:Microscopy, Confocal; D009363:Neoplasm Proteins / Q000032:analysis; D009930:Organic Chemicals; D009943:Organophosphorus Compounds; D011419:Propidium; D013194:Staining and Labeling / Q000379:methods*; D013347:Subcellular Fractions / Q000201:enzymology; D014407:Tumor Cells, Cultured
D002412:Cations; D004396:Coloring Agents; D005456:Fluorescent Dyes; D009363:Neoplasm Proteins; D009930:Organic Chemicals; D009943:Organophosphorus Compounds; C117141:fast red violet; C026282:naphthol AS-BI phosphate; D011419:Propidium; D008739:Methyl Green; D000135:Acid Phosphatase; D000165:Acridine Orange; D006416:Hematoxylin