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An improved microphotometry system for measurement of cytochrome P-450 in hepatocyte cytoplasm

J Watanabe and S Kanamura

Department of Anatomy, Kansai Medical University, Moriguchi, Osaka, Japan.

To measure cytochrome P-450 (P-450) content in hepatocyte cytoplasm, we developed a dual monochromator-equipped microphotometry system (KWSP- 1). Simultaneous measurements of absorbance at 450 and 490 nm with narrow band width (0.5 nm) and small spot size (2 microns) were accomplished by this system. Corresponding fields in serial sections could be easily and rapidly identified under the Nomarski imaging mode of KWSP-1. Photometric accuracy and repeatability of wavelength setting of KWSP-1 were also satisfactory for measurement of P-450. With this system, it is thus possible to measure the extinction of P-450 from many small measuring areas and to precisely determine P-450 content in the cytoplasm of rat hepatocytes. A microphotometric method was developed using cuvette slides and two serial 10-microns thick sections (mapping method). The intracellular distribution of P-450 in individual hepatocytes could be visualized by the mapping method with KWSP-1. However, this method was not applicable to tissue sections containing hemoglobin larger than 4 microM.

Volume 39, Issue 5, pp. 689-694, 05/01/1991
Copyright © 1991 by The Histochemical Society


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K. Mino, J. Watanabe, and S. Kanamura
Effect of 3-Methylcholanthrene Administration on Expression of Cytochrome P-450 Isoforms Induced by Phenobarbital in Rat Hepatocytes
J. Histochem. Cytochem., October 1, 1998; 46(10): 1151 - 1160.
[Abstract] [Full Text]



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