A method to compensate for light attenuation with depth, in three-dimensional DNA image cytometry using a confocal scanning laser microscope

A. Liljeborg, Magdalena Czader, A. Porwit

Research output: Contribution to journalArticle

25 Citations (Scopus)

Abstract

A method to compensate for attenuation of detected light with increased depth of the collected optical section, and its application in three-dimensional (3-D) DNA image cytometry is described. The method is based on studying the stack of 2-D histograms that can be formed from each consecutive pair of sections in a stack of optical serial sections. An attenuation factor is calculated interactively and a new compensated section series is computed. Formalin-fixed paraffin-embedded rat tissue was stained with propidium iodide. Each cell nucleus is extracted by thresholding and its total intensity is calculated. The coefficient of variation (CV) of the total intensity of all cells in each stack is computed. For comparison the CV of the same cells is computed in the uncompensated stacks. This study shows a significantly lower CV for the compensated data, thus contributing to the accuracy of DNA quantification in 3-D DNA image cytometry.

Original languageEnglish (US)
Pages (from-to)108-114
Number of pages7
JournalJournal of Microscopy
Volume177
Issue number2
StatePublished - 1995
Externally publishedYes

Fingerprint

Image Cytometry
cytometry
Three-Dimensional Imaging
Lasers
Microscopes
DNA
deoxyribonucleic acid
attenuation
microscopes
Scanning
Light
scanning
lasers
coefficients
Propidium
Cell Nucleus
Formaldehyde
Paraffin
Paraffins
Rats

Keywords

  • Compensation
  • Confocal
  • DNA image cytometry
  • Histostack
  • Light attenuation
  • Microscopy

ASJC Scopus subject areas

  • Instrumentation

Cite this

A method to compensate for light attenuation with depth, in three-dimensional DNA image cytometry using a confocal scanning laser microscope. / Liljeborg, A.; Czader, Magdalena; Porwit, A.

In: Journal of Microscopy, Vol. 177, No. 2, 1995, p. 108-114.

Research output: Contribution to journalArticle

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