Summary
A new method is described for staining cell nuclei with propidium iodide in preparations that have been processed for autoradiography. The permeability of the stripping film to the dye molecule has been studied, as have the staining conditions, in order to optimize the stoichiometry of the DNA-dye interaction. A procedure has also been set up to allow the simultaneous measurement of DNA content and grain count on the same cell.
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Allison, D. C., Mayne, J., Ridolpho, P. F., Bose, K., Chakerian, M. &Robertson, J. (1984) Computerized measurement of the DNA content, areas, and autoradiographic grains of the same nuclei: Demonstration that lightly [3H]Thymidine-labeled bone marrow cells are predominantly in G0/G1 and G2.J. Histochem. Cytochem. 32, 1197–203.
Alpen, E. L. &Johnston, M. E. (1967) DNA synthetic rate and DNA content of nucleated erythroid cells.Expl Cell Res. 47, 177–92.
Baserga, R. &Nemeroff, K. (1962) Two-emulsion radioautography.J. Histochem. Cytochem. 10, 628–35.
Burger, E. H. &Ploem, J. S. (1976) Quantitative autoradiography in combination with cytofluorometry for the study of sulphate incorporation in the developing epiphyseal plate.Proc. Kon. Ned. Akad. Wet. C 79, 9–25.
Crissman, H. A. &Steinkamp, J. A. (1973) Rapid, simultaneous measurement of DNA, protein and cell volume in single cells from large mammalian cell populations.J. Cell Biol. 59, 766–71.
Dörmer, P. (1973) Kinetics of erythropoietic cell proliferation in normal and anemic man. A new approach using quantitative14C-Autoradiography.Prog. Histochem. Cytochem. 6, 1–83.
Dörmer, P. (1981) Quantitative carbon-14 autoradiography at the cellular level: principles and application for cell kinetic studies.Histochem. J. 13, 161–71.
Dörmer, P., Pachmann, K. &Lau, B. (1978) Successive evaluation of multiple parameters in individual cells: the SEMPIC photometric system.Histochemistry 59, 17–27.
Dörmer, P. &Thiel, E. (1976) Methods of quantitative autoradiography using indicident light microphotometry.J. Histochem. Cytochem. 24, 145–51.
Freitas, M. I., Giordano, P. A. &Bottiroli, G. (1980) Improvement in microscope photometry by voltage to frequency convesion: analogue measurement and digital processing.J. Microsc. 124, 211–8.
Fujita, S. (1974) Analysis of cytokinetics by means of Feulgen cytofluorometry combined with3H-thymidine autoradiography.Expl Cell Res. 88, 395–401.
Fujita, S., Ashihara, T. &Fukuda, M. (1974) Simultaneous measurement of DNA content and grain count on an autoradiograph of Feulgen stained cells.Histochemistry 40, 155–63.
Giordano, P. A., Mazzini, G., Riccardi, A., Montecucco, C. M., Ucci, G. &Danova, M. (1984) Staining of cytoaudioradiographic preparations with propidium iodide.Abstracts VIIth Int. Congr. Histochem. Cytochem., Helsinki, Finland, p. 124.
Goldstein, D. J. &Williams, M. A. (1971) Quantitative autoradiography: an evaluation of visual grain counting, reflectance microscopy, gross absorbance measurments and flying-spot microdensitometry.J. Microsc. 94, 215–39.
Hughes, H. C., Meyer, P. M., Meyer, J. W., Meyer, D. R. &Bresnahan, J. C. (1977) An inexpensive microphotometer system for measuring silver grain densities in autoradiographs.Stain Technol. 52, 79–83.
Lipkin, L. E., Lemkin, P. &Carman, G. (1974) Automated autoradiographic grain counting in human determined context.J. Histochem. Cytochem. 22, 755–65.
Longoni, A., Cova, S., Giordano, P. A. &Freitas, I. (1985) A microspectrofluorometer system based on real time digital techniques.Analyt. Chem., in press.
Mazzini, G., Giordano, P., Montecucco, C. M. &Riccardi, A. (1980) A rapid cytofluorometric method for quantitative DNA determination on fixed smears.Histochem. J. 12, 153–68.
Pearse, A. G. E. (1980)Histochemistry: Theoretical and Applied. Vol. 1., 3rd edn, pp. 303–45 Edinburgh, London, New York: Churchill Livingstone.
Prensky, W. (1971) Automated image analysis in autoradiography.Expl Cell Res. 68, 388–94.
Rogers, A. W. (1973)Techniques of Auforadiography. 2nd edn. Amsterdam, London, New York: Elsevier.
Rüter, A., Aus, H. M., Harms, H., Haucke, M., Ter Maulen, V., Maurer-Schultze, B., Korr, H. &Kellerer, A. (1979) Automated quantitative analysis of single and double label autoradiographys.J. Histochem. Cytochem. 27, 217–24.
Schuette, W. H., Chen, S. S., Occhipinti, S. J., Mujagic, H. S. &Shackney, S. E. (1983) Automated radioautographic grain counting. Correction for grain overlap.Cell Tiss. Kinet. 16, 221–7.
Sklarew, R. J. (1982) Simultaneous Feulgen densitometry and autoradiographic grain counting with the quantimet 720D image-analysis system. I. Estimation of nuclear DNA content in3H-Thymidinelabeled cells.J. Histochem. Cytochem. 30, 35–48.
Sklarew, R. J. (1984) Cytokinetics of subpopulations in mixed heteroploid tumors by television imaging. I. Deconvolution of the S-phase DNA ploidy composition. II. Analysis of the S-phase emptying profile of ploidy subpopulations.J. Histochem. Cytochem. 32, 413–20.
Thurston, J. M. &Joftes, D. L. (1963) Stains compatible with dipping radioautography.Stain Technol. 38, 231–5.
Vendrely, C. (1971) Cytophotometry and histochemistry of the cell cycle. InThe Cell Cycle and Cancer (edited byBaserga, R.), pp. 227–68 New York: Marcel Dekker.
Wachsmuth, E. D., Becker, M., Grieve, A. P. &Maurer-Schultze, B. (1984) Fully automated and fast image analysis of autoradiographys with a TAS-Leitz. Determination of size, Feulgen fluorescence and grain counts of individual nuclei and their evaluation by a simplified cluster analysis.Histochemistry 81, 227–36.
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Giordano, P.A., Mazzini, G., Riccardi, A. et al. Propidium iodide staining of cytoautoradiographic preparations for the simultaneous determination of DNA content and grain count. Histochem J 17, 1259–1270 (1985). https://doi.org/10.1007/BF01002507
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DOI: https://doi.org/10.1007/BF01002507