Increased accuracy and speed of absorption cytometric DNA measurements by automatic corrections for nuclear darkness.

Autor: Allison DC, Lawrence GN, Ridolpho PF, O'Grady BJ, Rasch RW, Rasch EM
Jazyk: angličtina
Zdroj: Cytometry [Cytometry] 1984 May; Vol. 5 (3), pp. 217-27.
DOI: 10.1002/cyto.990050302
Abstrakt: We have developed a method of calculating the average local absorbance (ALA) of a nucleus from the integrated nuclear absorbance and area. One can use the ALA, along with nuclear areas measured at different point absorbance thresholds, to determine whether a nucleus is stained too lightly or too darkly for accurate absorption measurements; this allows selection of an optimal light wavelength for the performance of these measurements. The ALA can also be used for automatic and instantaneous correction of integrated absorbance values from darkly stained cells. This allows the rapid measurement of the integrated absorbances of a large number of nuclei that are heterogeneous in stain intensity. Coefficients of variation of approximately 3% are obtained for the integrated absorbances of nuclei of nontransformed G0/G1 cells. This correction method can be applied with any image densitometer that generates both integrated absorbance and area values.
Databáze: MEDLINE