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Normal and perturbed Chinese hamster ovary cells: correlation of DNA, RNA, and protein content by flow cytometry

Quantitative, correlated determinations of DNA, RNA, and protein, as well as RNA to DNA and RNA to protein ratios, were performed on three- color stained cells using a multiwavelength-excitation flow cytometer. DNA-bound Hoechst 33342 (blue), protein-fluorescein isothiocyanate (green), and RNA-bound...

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Formato: Texto
Lenguaje:English
Publicado: The Rockefeller University Press 1985
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2113613/
https://www.ncbi.nlm.nih.gov/pubmed/2409095
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collection PubMed
description Quantitative, correlated determinations of DNA, RNA, and protein, as well as RNA to DNA and RNA to protein ratios, were performed on three- color stained cells using a multiwavelength-excitation flow cytometer. DNA-bound Hoechst 33342 (blue), protein-fluorescein isothiocyanate (green), and RNA-bound pyronin Y (red) fluorescence measurements were correlated as each stained cell intersected three spatially separated laser beams. The analytical scheme provided sensitive and accurate fluorescence determinations by minimizing the effects of overlap in the spectral characteristics of the three dyes. Computer analysis was used to generate two-parameter contour density profiles as well as to obtain numerical data for subpopulations delineated on the basis of cellular DNA content. Such determinations allowed for analysis of RNA to DNA and RNA to protein ratios for cells within particular regions of the cell cycle. The technique was used to study the interrelationship of DNA, RNA, and protein contents in exponentially growing Chinese hamster ovary cells as well as in cell populations progressing the cell cycle after release from arrest in G1 phase. The sensitivity of the method for early detection of conditions of unbalanced growth is demonstrated in the comparison of the differential effects of the cycle-perturbing agent, adriamycin, on cells treated either during exponential growth or while reversibly arrested in G1 phase.
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spelling pubmed-21136132008-05-01 Normal and perturbed Chinese hamster ovary cells: correlation of DNA, RNA, and protein content by flow cytometry J Cell Biol Articles Quantitative, correlated determinations of DNA, RNA, and protein, as well as RNA to DNA and RNA to protein ratios, were performed on three- color stained cells using a multiwavelength-excitation flow cytometer. DNA-bound Hoechst 33342 (blue), protein-fluorescein isothiocyanate (green), and RNA-bound pyronin Y (red) fluorescence measurements were correlated as each stained cell intersected three spatially separated laser beams. The analytical scheme provided sensitive and accurate fluorescence determinations by minimizing the effects of overlap in the spectral characteristics of the three dyes. Computer analysis was used to generate two-parameter contour density profiles as well as to obtain numerical data for subpopulations delineated on the basis of cellular DNA content. Such determinations allowed for analysis of RNA to DNA and RNA to protein ratios for cells within particular regions of the cell cycle. The technique was used to study the interrelationship of DNA, RNA, and protein contents in exponentially growing Chinese hamster ovary cells as well as in cell populations progressing the cell cycle after release from arrest in G1 phase. The sensitivity of the method for early detection of conditions of unbalanced growth is demonstrated in the comparison of the differential effects of the cycle-perturbing agent, adriamycin, on cells treated either during exponential growth or while reversibly arrested in G1 phase. The Rockefeller University Press 1985-07-01 /pmc/articles/PMC2113613/ /pubmed/2409095 Text en This article is distributed under the terms of an Attribution–Noncommercial–Share Alike–No Mirror Sites license for the first six months after the publication date (see http://www.rupress.org/terms). After six months it is available under a Creative Commons License (Attribution–Noncommercial–Share Alike 4.0 Unported license, as described at http://creativecommons.org/licenses/by-nc-sa/4.0/).
spellingShingle Articles
Normal and perturbed Chinese hamster ovary cells: correlation of DNA, RNA, and protein content by flow cytometry
title Normal and perturbed Chinese hamster ovary cells: correlation of DNA, RNA, and protein content by flow cytometry
title_full Normal and perturbed Chinese hamster ovary cells: correlation of DNA, RNA, and protein content by flow cytometry
title_fullStr Normal and perturbed Chinese hamster ovary cells: correlation of DNA, RNA, and protein content by flow cytometry
title_full_unstemmed Normal and perturbed Chinese hamster ovary cells: correlation of DNA, RNA, and protein content by flow cytometry
title_short Normal and perturbed Chinese hamster ovary cells: correlation of DNA, RNA, and protein content by flow cytometry
title_sort normal and perturbed chinese hamster ovary cells: correlation of dna, rna, and protein content by flow cytometry
topic Articles
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2113613/
https://www.ncbi.nlm.nih.gov/pubmed/2409095