Relation between fluorescent intensity and extinction in quantitative microfluorometry.
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Biomedical subjects
Publications and source records attributed to G Boeken.
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Methods in use for the choice of the two-wavelengths and the effect of a faulty choice of the two wavelengths in the two-wavelength cytophotometric method of Patau and Ornstein are studied by means of a computer simulation and in practice upon pararosaniline-Schiff stained nuclei. A simplification of the measuring strategy is proposed without affecting the accuracy.
Factors influencing the calculation of the relative amount of chromophore and the chromophore area by the two-wavelength method are examined. The study was carried out with the help of models and further tested on Feulgen stained preparations. Except for certain restrictions the difference between the chromophore area as calculated from the two transmissions measurements and the chromophore area obtained by planimetry can be used as a guide for determining the proper measuring conditions, including the choise of the two wavelengths.
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Three methods for the evaluation of the relative amount of DNA per nucleus by absorption cytophotometry are compared. A combination of the two-wavelength method (Patau, 1952; Ornstein, 1952), the one-wavelength two-area method (Garcia, 1965) and the determination of transmission through nuclear plugs is proposed in order to estimate systematic errors made by absorption cytophotometry.