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Standardization of tritium-sensitive film for quantitative autoradiography

DG Baskin, PE Filuk and WL Stahl

Veterans Administration Medical Center, Seattle, Washington 98108.

A method for the calibration of plastic tritium standards for use with LKB Ultrofilm is described and validated. This method uses 12-microns cryostat slices of frozen liver which have been labeled with [3H]- formaldehyde and extracted with chloroform-methanol to remove lipids. Quantitative autoradiographic measurement of 3H radioactivity in the liver slices was underestimated by 35% when lipids were not extracted. Plastic sections impregnated with tritium were calibrated in terms of lipid-extracted, tissue-equivalent radioactivity content. Calibrated standard curves for these plastic standards were closely fit (p = 0.99) by second order polynomial equations for exposures of 1, 4, 7, 13, 28, and 104 days. The equations are generally useful for any plastic tritium standards.

Volume 37, Issue 9, pp. 1337-1344, 09/01/1989
Copyright © 1989 by The Histochemical Society


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