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

David C. Allison, George N. Lawrence, Paul F. Ridolpho, Brian J. O'Grady, Robert W. Rasch, Ellen M. Rasch

Research output: Contribution to journalArticle

Abstract

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.

Original languageEnglish (US)
Pages (from-to)217-227
Number of pages11
JournalCytometry
Volume5
Issue number3
DOIs
StatePublished - May 1984

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Keywords

  • Absorption cytometry
  • Feulgen stain
  • measurement of cellular DNA

ASJC Scopus subject areas

  • Pathology and Forensic Medicine
  • Biophysics
  • Hematology
  • Endocrinology
  • Cell Biology

Cite this

Allison, D. C., Lawrence, G. N., Ridolpho, P. F., O'Grady, B. J., Rasch, R. W., & Rasch, E. M. (1984). Increased accuracy and speed of absorption cytometric DNA measurements by automatic corrections for nuclear darkness. Cytometry, 5(3), 217-227. https://doi.org/10.1002/cyto.990050302