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British Journal of Cancer logoLink to British Journal of Cancer
. 1977 Nov;36(5):592–600. doi: 10.1038/bjc.1977.236

Fluorescence discrimination between diploid cells on their RNA content: a possible distinction between clonogenic and non-clonogenic cells.

J V Watson, S H Chambers
PMCID: PMC2025403  PMID: 588421

Abstract

Flow cytofluorimetric techniques, using acridine orange fluorescence to measure RNA and DNA simultaneously in EMT6 cells, have been employed to discriminate between three diploid DNA populations in vivo on the basis of their RNA content. Cells with the lowest RNA levels seem to be in the process of disintegration. Cells with the highest RNA levels correspond to those with the highest plating efficiency, and those with intermediate RNA levels are those with the lowest plating efficiency. In vitro studies have shown that log-phase cells have higher RNA levels than cells in the late plateau phase of growth.

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Selected References

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  1. ARMSTRONG J. A. Histochemical differentiation of nucleic acids by means of induced fluorescence. Exp Cell Res. 1956 Dec;11(3):640–643. doi: 10.1016/0014-4827(56)90173-2. [DOI] [PubMed] [Google Scholar]
  2. BICKIS I., VON BERTALANFFY L. Identification of cytoplasmic basophilia (ribonucleic acid) by fluorescence microscopy. J Histochem Cytochem. 1956 Sep;4(5):481–493. doi: 10.1177/4.5.481. [DOI] [PubMed] [Google Scholar]
  3. Bradley D. F., Wolf M. K. AGGREGATION OF DYES BOUND TO POLYANIONS. Proc Natl Acad Sci U S A. 1959 Jul;45(7):944–952. doi: 10.1073/pnas.45.7.944. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Darzynkiewicz Z., Traganos F., Sharpless T., Melamed M. R. Conformation of RNA in situ as studied by acridine orange staining and automated cytofluorometry. Exp Cell Res. 1975 Oct 1;95(1):143–153. doi: 10.1016/0014-4827(75)90619-9. [DOI] [PubMed] [Google Scholar]
  5. Darzynkiewicz Z., Traganos F., Sharpless T., Melamed M. Thermally-induced changes in chromatin of isolated nuclei and of intact cells as revealed by acridine orange staining. Biochem Biophys Res Commun. 1974 Jul 10;59(1):392–399. doi: 10.1016/s0006-291x(74)80219-6. [DOI] [PubMed] [Google Scholar]
  6. Gersch N. F., Jordan D. O. Interaction of DNA with aminoacridines. J Mol Biol. 1965 Aug;13(1):138–156. doi: 10.1016/s0022-2836(65)80085-7. [DOI] [PubMed] [Google Scholar]
  7. Krishan A. Rapid flow cytofluorometric analysis of mammalian cell cycle by propidium iodide staining. J Cell Biol. 1975 Jul;66(1):188–193. doi: 10.1083/jcb.66.1.188. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Traganos F., Darzyndiewicz Z., Sharpless T., Melamed M. R. Denaturation of deoxyribonucleic acid in situ effect of formaldehyde. J Histochem Cytochem. 1975 Jun;23(6):431–438. doi: 10.1177/23.6.239052. [DOI] [PubMed] [Google Scholar]
  9. Traganos F., Darzynkiewicz Z., Sharpless T., Melamed M. R. Simultaneous staining of ribonucleic and deoxyribonucleic acids in unfixed cells using acridine orange in a flow cytofluorometric system. J Histochem Cytochem. 1977 Jan;25(1):46–56. doi: 10.1177/25.1.64567. [DOI] [PubMed] [Google Scholar]
  10. Twentyman P. R., Bleehen N. M. The sensitivity to bleomycin of a solid mouse tumour at different stages of growth. Br J Cancer. 1974 Nov;30(5):469–472. doi: 10.1038/bjc.1974.222. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Twentyman P. R., Watson J. V., Bleehen N. M., Rowles P. M. Changes in cell proliferation kinetics occurring during the life history of monolayer cultures of a mouse tumour cell line. Cell Tissue Kinet. 1975 Jan;8(1):41–50. doi: 10.1111/j.1365-2184.1975.tb01205.x. [DOI] [PubMed] [Google Scholar]
  12. Twentyman P. R., Watson J. V. Separation of clonogenic tumour cells from EMT6 mouse mammary tumours. Br J Cancer. 1977 Jan;35(1):120–122. doi: 10.1038/bjc.1977.12. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Watson J. V. The application of age distribution theory in the analysis of cytofluorimetric DNA histogram data. Cell Tissue Kinet. 1977 Mar;10(2):157–169. doi: 10.1111/j.1365-2184.1977.tb00141.x. [DOI] [PubMed] [Google Scholar]

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