A comparative study of quantitative stains for DNA in image cytometry
1Department of Cellular Pathology, Armed Forces Institute of Pathology, Washington, D.C.
Analytical and Quantitative Cytology and Histology
|August 11, 1991
Summary
This study evaluated DNA stains for image cytometry, finding Feulgen staining, particularly acriflavine-Feulgen, offers the most reproducible and accurate nuclear DNA measurements. Carnoy
Area of Science:
- Cell Biology
- Histology
- Biotechnology
Background:
- Accurate nuclear DNA measurement is crucial for image cytometry.
- Reproducibility of DNA stains impacts diagnostic reliability.
Purpose of the Study:
- To assess the reproducibility of various nuclear DNA stains for image cytometry.
- To identify optimal staining methods for quantitative DNA analysis.
Main Methods:
- Tested seven DNA stains (four Feulgen, gallocyanine-chromalum, acridine orange, propidium iodide) on mouse and rat tissues.
- Utilized video image analysis and scanning microspectrophotometry for measurements.
- Performed nested analysis of variance to quantify variability sources.
Main Results:
- All tested stains were quantitative and reproducible for DNA measurement.
- Absorbance measurements showed lower coefficients of variation (CV) than fluorescence.
- Acriflavine-Feulgen staining with Carnoy's fixation yielded minimal variation and accurate ploidy ratios.
Conclusions:
- Feulgen staining is the most effective technique for DNA analysis via image cytometry.
- Acriflavine-Feulgen staining with Carnoy's fixation is recommended for optimal results.
- This method provides superior accuracy and reproducibility compared to other tested stains.
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