Advertisement

The Histochemical Journal

, Volume 6, Issue 3, pp 259–278 | Cite as

Methodological and instrumentational aspects of cytofluorometry

  • G. Prenna
  • G. Mazzini
  • S. Cova
Papers

Synopsis

Some aspects of cellular fluorescence phenomena are examined for the purpose of establishing the experimental conditions required in order to obtain reliable data by cytofluorometric measurements. Limitations of the acceptable range of extinction values are established, and devices to operate within this range are suggested. The inner filter effect and its associated errors are considered, together with methods to avoid them. Quenching effects are also outlined. The photodecomposition phenomenon is dealt with, and the operational devices to limit its influence are illustrated. The influence on the measured values of autofluorescence and of other background sources is examined. Some aspects of electronic instrumentation suitable for accurate and precise measurement of light intensities, even at low levels, are also discussed.

Keywords

Light Intensity Reliable Data Precise Measurement Acceptable Range Operational Device 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

Preview

Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.

References

  1. Böhm, N. &Sprenger, E. (1968). Fluorescence cytophotometry: a valuable method for the quantitative determination of nuclear Feulgen-DNA.Histochemie. 16, 100–18.Google Scholar
  2. Cova, S., Prenna, G. &Mazzini, G. (1972). Microspectrofluorometry by single photon multiscaling.Abstracts of 4th International Congress of Histochemistry and Cytochemistry, Kyoto. pp. 161–2.Google Scholar
  3. Cova, S., Bertolaccini, M. &Bussolatic, C. (1973). The measurement of luminescence waveforms by single photon techniques.Phys. Stat. Sol. (a) 18, 11–62.Google Scholar
  4. Cova, S., Prenna, G. &Mazzini, G. (1974). Digital microspectrofluorimetry by multichannel scaling and single photon detection.Histochem. J 6, 279–99.Google Scholar
  5. Deeley, E. M. (1955). An integrating microdensitometer for biological cells.J. Sci. Instrum. 32, 263–7.Google Scholar
  6. Förster, T. (1959). Transfer mechanisms of electronic excitation.Discuss. Faraday Soc. 27, 7–17.Google Scholar
  7. Göhde, W. &Dittrich, W. (1970). Simultane Impulsfluorimetrie des DNS- und Proteingehaltes von Tumorzellen.Z. Anal. Chem. 252, 328–30.Google Scholar
  8. Kamentsky, L. A. (1970). The rapid cell spectrophotometer. In:Automated Cell Identification and Cell Sorting (eds. G. L. Wied & G. F. Bahr), pp. 111–21. New York and London: Academic Press.Google Scholar
  9. Perrin, F. (1923). Observations sur la fluorescence.C. r. hebd. Séanc. Acad. Sci. Paris. 177, 469–75.Google Scholar
  10. Prenna, G. &Zanotti, L. (1964). A new cytofluorometric method for quantitative determinations of DNA.Abstracts of 2nd International Congress of Histochemistry and Cytochemistry. Frankfurt/Main. p. 231.Google Scholar
  11. Prenna, G. (1964). Caratteristiche del [2'-(p-aminofenil)-6-metil-2, 6′-bibenzotiazolo]-SO2, un nuovo reagente tipo Schiff fluorescente and elevata sensibilità.Riv. Istoch. norm. pat. 10, 469–74.Google Scholar
  12. Prenna, G. &Bianchi, U. A. (1964). Reazioni di Feulgen fluorescenti e loro possibilità citofluorometriche quantitative. 5) Citofotometria quantitativa in fluorescenza e in assorbimento della reazione di Feulgen eseguita con acriflavina-SO2.Riv. Istoch. norm. pat. 10, 667–76.Google Scholar
  13. Prenna, G. (1968). Qualitative and quantitative applications of fluorescent Schiff-type reagents.Mikroskopie. 23, 150–4.Google Scholar
  14. Prenna, G., Leiva, S. &Mazzini, G. (1972). Quantitative DNA cytofluorometry by the conventional Feulgen reaction.Abstracts of 4th International Congress of Histochemistry and Cytochemistry, Kyoto, pp. 201–2.Google Scholar
  15. Prenna, G. (1973). Fluorescent Schiff-type reagents. In:The Encyclopedia of Microscopy and Microtechnique (ed. P. Gray), pp. 186–9. London: Van Nostrand-Reinhold.Google Scholar
  16. Prenna, G., Leiva, S. &Mazzini, G. (1974). Quantitative DNA cytofluorometry by the conventional Feulgen reaction.Histochem. J. 6, in press.Google Scholar
  17. Rigler, R. (1966). Microfluorometric characterization of intracellular nucleic acids and nucleoproteins by acridine orange.Acta physiol. scand. 67, Suppl. 267, 1–122.Google Scholar
  18. Ritzén, M. (1967). Cytochemical identification and quantitation of biogenic monoamines. A microspectrofluorometric and autoradiographic study.M. D. Thesis, Stockholm.Google Scholar
  19. Rosanov, Ju. M. &Kudryavtsev, B. N. (1967). A method of fluorescence cytophotometry for the purpose of a quantitative determination of DNA (Russ).Tsitologiya. 9, 361–7.Google Scholar
  20. Ruch, F. (1966). Determination of DNA content by microfluorometry. In:Introduction to Quantitative Cytochemistry, Vol. I (ed. G. L. Wied), pp. 281–94. New York and London: Academic Press.Google Scholar
  21. Ruch, F. (1970). Principles and some applications of cytofluorometry. In:Introduction to Quantitative Cytochemistry, Vol. II (eds. G. L. Wied & G. F. Bahr), pp. 431–50. New York and London: Academic Press.Google Scholar
  22. Sacchi, C. A., Svelto, O. &Prenna, G. (1974). Pulsed Tunable Lasers in cytofluorometry.Histochem. J. 6, 251–8.Google Scholar
  23. Thaer, A. A. (1966). Instrumentation for microfluorometry. In:Introduction to Quantitative Cytochemistry, Vol. I (ed. G. L. Wied), pp. 409–26. New York and London: Academic Press.Google Scholar

Copyright information

© Chapman and Hall Ltd 1974

Authors and Affiliations

  • G. Prenna
    • 1
  • G. Mazzini
    • 1
  • S. Cova
    • 2
  1. 1.Centro di Studio per l'Istochimica del C.N.R., Istituto di Anatomia ComparataUniversitá di PaviaItaly
  2. 2.Istituto di Fisica del Politecnica di MilanoItaly

Personalised recommendations