Synopsis
Edström's method for the ultramicrochemical determination of RNA and DNA in individual cells was modified for the measurement of extinction in u.v. light with the aid of the Zeiss scanning microspectrophotometer UMSP-I. With this new procedure, nucleic acids down to about 3 pg RNA or about 4 pg DNA can be measured with a very high accuracy.
The method was applied to enzymatically isolated rat liver parenchymal cells. A mean DNA content of 6.52 pg was found for diploid cells. The DNA content of mononuclear cells of different ploidy levels and of binuclear cells showed a close proportionality with the nuclear ploidy and the number of nuclei per cell. The RNA content of mononuclear diploid cells amounted to 33.4 pg, yielding an RNA/DNA ratio of 5.12. The RNA/DNA ratio was similar for binuclear and mononuclear cells of the same ploidy level but decreased considerably with increasing nuclear ploidy.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Berry, M. N. &Friend, D. S. (1969). High-yield, preparation of isolated rat liver parenchymal cells.J. Cell. Biol. 43, 506–20.
Brattgård, S.-O., Edström, J.-E. &Hydén., H. (1957). The chemical changes in regenerating neurons.J. Neurochem. 1, 316–25.
Brattgård, S.-O., Edström, J.-E. &Hydén, H. (1958). The reproductive capacity of the neuron in retrograde reaction.Expl. Cell. Res. (suppl.).5, 185–200.
De Fonbrune, P. (1949).Techniques de Manipulation, pp. 121–57. Monographs de l'Institut Pasteur, Masson Cie Ed., Paris.
Edström, J.-E. (1953). Ribonucleic acid mass and concentration in individual nerve cells. A new method for quantitative determination.Biochim. biophys. Acta 12, 361–86.
Edström, J.-E. (1956). The content and the concentration of ribonucleic acid in motor anterior horn cells from the rabbit.J. Neurochem. I, 159–65.
Edström, J.-E. (1959). Ribonucleic acid changes in the motoneurons of the frog during axon regeneration.J. Neurochem. 5, 43–9.
Edström, J.-E. (1964). Microextraction and microelectrophoresis for determination and analysis of nucleic acids in isolated cellular units. In:Methods in Cell Physiology. Vol I. (ed. D. M. Prescott), pp. 417–47.
Edström, J.-E. &Eichner, D. (1957). Quantitative Untersuchungen über den Ribonukleinsaüregehalt der Netzhautganglienzellen bei Rind und Mensch.Z. mikrosk.-anat. Forsch. 63, 413–21.
Edström, J.-E. &Eichner, D. (1958). Quantitative Ribonukleinsaüre-Untersuchungen an den Ganglienzellen des Nucleus supraopticus der Albino-Ratte unter experimentellen Bedingungen (Kochsalz-Belastung).Z. Zellforsch. 48, 187–200.
Edström, J.-E. &Kawiak, J. (1961). Microchemical deoxyribonucleic acid determination in individual cells.J. biophys. biochem. Cytol 9, 619–26
Edström, J.-E. &Neuhoff, V. (1973). Micro-electrophoresis for RNA and DNA base analysis. In:Micromethods in Molecular Biology (ed. V. Neuhoff), pp. 213–56. Berlin: Springer Verlag.
Edström, J.-E. &Pigon, A. (1958). Relation between surface, ribonucleic acid content and nuclear volume in encapsulated spinal ganglion cells.J. Neurochem. 3, 95–9.
Epstein, C. J. (1967). Cell size, nuclear content, and the development of polyploidy in the mammalian liver.Proc. natn. Acad. Sci. 57, 327–34.
Hallén, O., Edström, J.-E. &Hamberger, A. (1965). Cytochemical response to acoustic stimuli in the spiral ganglion cells of guinea pigs.Acta oto-laryng. 60, 121–8.
James, J. (1965). Constancy of nuclear DNA and accuracy of cytophotometric measurement.Cytogenetics 4, 19–27.
James, J. (1968). Feulgen-DNA changes in rat liver cell nuclei during the early phase of ischaemic necrosis.Histochemie 13, 312–22.
James, J. (1973). Extinction coefficients in Feulgen-DNA scanning photometry of human lymphocytes.Acta Cytol. 17, 15–18.
James, J. (1976).Light Microscopic Techniques in Biology and Medicine, pp. 186–7., Leiden: Stenfert Kroese.
Kiefer, G. (1970). Recent developments in gallocyanine-chrome alum staining. In:Introduction to Quantitative Cytochemistry. Vol. II (eds. G. L. Wied & G. F. Bahr), pp. 199–208. New York, London: Academic Press.
Morselt, A. F. W. &Van Wijgerden, H. G. M. (1975). Microphotometry of rat liver nucleoproteins during the cell cycle, and comparison of diploid nuclei in the G2 period with tetraploid nuclei.Histochemistry 44, 87–93.
Rouser, G., Fleischer, S. &Yamamoto, A. (1970). Two dimensional, thin layer chromotographic separation of polar lipids and determination of phospholipids by phosphorus analysis of spots.Lipids 5, 494–6.
Schiemer, A. G. (1969) Über Schnellphotometrie mit dem Zeiss'schen Universalmikrospektrophotometer (UMSP-I).Z. wiss. Mikrosk. 69, 244–56.
Tas, J., Oud, P. &James, J. (1974). The naphthol yellow S stain for proteins tested in a model system of polyacrylamide films and evaluated for practical use in histochemistry.Histochemistry 40, 231–40.
Vendrely, R. (1955). The deoxyribonucleic acid content of the nucleus. In:The Nucleic Acids, Chemistry and Biology. Vol. II. (eds. E. Chargaff & J. N. Davidson), pp. 155–92. New York: Academic Press.
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Roozemond, R.C. Ultramicrochemical determination of nucleic acids in individual cells using the Zeiss UMSP-I microspectrophotometer. Application to isolated rat hepatocytes of different ploidy classes. Histochem J 8, 625–638 (1976). https://doi.org/10.1007/BF01003963
Received:
Revised:
Issue Date:
DOI: https://doi.org/10.1007/BF01003963