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A number of histochemical chromogenic substrates for alkaline phosphatase are commercially available and give reaction products with a range of colours for brightfield examination. Some of these reaction products are also fluorescent, exhibiting a wide excitation range and a broad emission peak. We report here that one of these substrates, Vector Blue III, yields a stable, strongly fluorescent reaction product with an excitation peak around 500 nm and a large Stokes shift to an emission peak at 680 nm. The reaction product can be excited using a mercury lamp with a fluorescein excitation filter or an argon ion laser at 488 nm or 568 nm, and the emission detected using a long-pass filter designed for Cy-5. Thus, a single substrate is suitable for brightfield imaging of tissue sections and high-resolution analysis of subcellular detail, using a confocal laser scanning microscope, in the same specimen.

Type

Journal article

Journal

Histochem J

Publication Date

08/1998

Volume

30

Pages

577 - 581

Keywords

Adrenocorticotropic Hormone, Alkaline Phosphatase, Animals, Cell Line, Enzyme-Linked Immunosorbent Assay, Fluorescence, Immunoenzyme Techniques, Microscopy, Confocal, Microscopy, Fluorescence, Mitochondria, Pituitary Gland, Rats, Spectrometry, Fluorescence