Localization of tissue antigens by enzyme immunocytochemistry

Specific cytochemical procedures for staining enzymes are available for both light and electron microscopy. At the light microscope level, mainly peroxidase but also alkaline phosphatase and occasionally glucose oxidase are used as labels. Because cytochemical staining procedures for enzymes give rise to reaction products of different colors, enzyme-labeled antibodies can be used for the simultaneous detection of two different cellular constituents. Alternatively, paired staining techniques combining autoradiography or immunofluorescence and immunoenzymatic staining are employed for the detection of two antigens. Detection of antigens by immunoenzymatic and immunofluorescence staining give comparable results both in localization and sensitivity.

At the electron microscope level, peroxidase is almost exclusively used as the label because 1) its cytochemical staining permits precise localization and identification of ultrastructural details, and 2) relatively low molecular weight peroxidase-antibody conjugates penetrate more easily into the interior of fixed cells.

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