Studies in histochemistry. XLVIII. Dry mass of mast cells measured by interference microscopy and x-ray absorption.
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A method for the determination of the DNA content of isolated nuclei of different ploidy has been developed. It is based on measurement of the nuclear dry mass, with an integrating microinterferometer, before and after DNase treatment. The values found are slightly low, because, as indicated by biochemical determinations, consistently 5% to 8% of DNA is not extracted by DNase under these conditions. The average DNA values thus obtained for diploid and tetraploid nuclei of adult rat liver are 7.7 and 15.6 pg (10(-12) g), respectively. Definite advantages of this procedure are: i) comparisons with biochemical determinations to give DNA values for each class of ploidy, ii) comparisons with histophotometry of the Feulgen dye-DNA complex to give absolute values instead of arbitrary units.
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Cells, obtained from human leg by successive treatments with trypsin, were air dried on microscope slides before mounting in glycerol. Dry weights and projected areas of individual cells were measured using a Vickers M 86 scanning microinterferometer. The dry weights of cells varied from 100 pg for basal cells to 700 pg for large squames. Corresponding projected areas varied from 100 to 1500 mum2.