Summary
Deparaffinized and Feulgen-stained sagittal sections of the mouse brain were studied interferometrically in order to measure optical path differences of euchromatin and heterochromatin of various cell types. Furthermore, the ratio eu-: heterochromatin of each cell type was determined. From these data mass densities of karyoplasm and, finally, correction factors of 3H-β-self-absorption were calculated for comparing grain numbers of different cell types in quantitative autoradiographic studies after application of tritium-labelled substances. Remarkable differences of correction factors up to a factor of 2.18 were found. Furthermore, the actual section thickness was determined interferometrically. A reduction to about 0.60 of the microtome setting was measured in two different areas of the brain. Using mass densities together with actual section thickness correction factors for a thickness of 1 μm were calculated. This was done also for cell types outside the brain the data of which were taken from literature. Thus, differences in correction factors up to about a factor of 4 were found pointing out the importance of considering 3H-β-self-absorption in quantitative autoradiographic studies.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Ada GL, Humphrey JH, Askonas BA, McDevitt HO, Nossal GJV (1966) Correlation of grain counts with radioactivity (125I and tritium) in autoradiography. Exp Cell Res 41:557–572
Alexander GM, Schwartzman RJ, Bell RD, Yu J, Renthal A (1981) Quantitative measurement of local cerebral metabolic rate for glucose utilizing tritiated 2-deoxyglucose. Brain Res 223:59–67
Anthony A, Colurso GJ, Bocan TMA, Doebler JA (1984) Interferometric analysis of intrasection and intersection thickness variability associated with cryostat microtomy. Histochem J 16:61–70
Arnold G (1965) An autoradiographic study of RNA synthesis in isolated salivary glands of Drosophila hydei. I. Autoradiographic studies. J Morphol 116:65–88
Bencke G (1966) Application of interference microscopy to biological material. In: Wied GL (ed) Introduction to quantitative cytochemistry. Academic Press, New York London, pp 63–92
Cavanagh JB, Lewis PD (1969) Perfusion-fixation, colchicine and mitotic activity in the adult rat brain. J Anat 104:341–350
Caviness VS, Barkley DS (1971) Section thickness and grain count variation in tritium autoradiography. Stain Technol 46:131–135
D'Ans J, Lax E (1949) Taschenbuch für Chemiker und Physiker, 2. Aufl. Springer, Berlin Göttingen Heidelberg
Davies HG (1958) The determination of mass and concentration by microscope interferometry. In: Danielli JF (ed) General cytochemical methods, Vol 1. Academic Press, New York, pp 55–161
Dörmer P, Brinkmann W (1972) Quantitative 14C-Autoradiographie einzelner Zellen. Histochemie 29:248–264
Falk GJ, King RC (1963) Radioautographic efficiency for tritium as a function of section thickness. Radiat Res 20:466–470
Fitzgerald PJ, Eidinoff ML, Knoll JE, Simmel EB (1951) Tritium in radiography. Science 114:494–498
Franca LCM, Foraker AG (1960) Interferometric study of astrocytes in gliosis and astrocytoma. Arch Neurol 3:416–419
Herkenham M, Sokoloff L (1984) Quantitative receptor autoradiography: tissue defatting eliminates differential selfabsorption of tritium radiation in gray and white matter of brain. Brain Res 321:363–368
Korr H (1981) Light microscopical autoradiography of nervous tissue. In: Heym C, Forssmann WG (eds) Techniques in neuroanatomical research. Springer, Berlin Heidelberg New York, pp 218–244
Korr H, Schultze B (1985) Unscheduled DNA synthesis (UDS) in glial and endothelial cells of the untreated mouse brain. Cell Tissue Kinet (in press)
Landolt H, Börnstein R (1912) Physikalisch-chemische Tabellen. 4. Aufl. Börnstein R, Roth WA (eds). Springer, Berlin
Ling EA, Leblond CP (1973) Investigation of glial cells in semithin sections. II. Variation with age in the numbers of the various glial cell types in rat cortex and corpus callosum. J Comp Neurol 149:73–82
Mares V, Schultze B, Maurer W (1974) Stability of DNA in Purkinje cell nuclei of the mouse. An autoradiographic study. J Cell Biol 63:665–674
Maurer W, Primbsch E (1964) Größe der β-Selbstabsorption bei der 3H-Autoradiographie. Exp Cell Res 33:8–18
Maurer W, Schultze B (1969) Problems in autoradiographic studies of DNA, RNA, and protein synthesis. In: Roth LJ, Stumpf WE (es) Autoradiography of diffusible substances. Academic Press, New York London, pp 15–28
Pelc SR, Welton MGE (1967) Quantitative evaluation of tritium in autoradiography and biochemistry. Nature 216:925–927
Perry RP, Errera M, Hell A, Dürwald H (1961) Kinetics of nucleoside incorporation into nuclear and cytoplasmic RNA. J Biophys Biochem Cytol 11:1–13
Pollister AW (1965) An autoradiographic study of RNA synthesis in isolated salivary glands of Drosophila hydei. II. Interferometric studies. J Morphol 116:89–98
Roger AW (1979) Techniques of autoradiography. 3rd edn. Elsevier/North-Holland Biomedical Press, Amsterdam New York Oxford
Vöhringer A, Maurer W (1971) Messung der Spezifischen Optischen Streifenverschiebung bei interferometrischen Massenbestimmungen von fixiertem Eiweiß und Nukleoprotein. Histochemie 25:234–246
Author information
Authors and Affiliations
Additional information
Dedicated to the sixtieth birthday of Prof. Dr. Brigitte Maurer-Schultze, Würzburg.
Rights and permissions
About this article
Cite this article
Korr, H. Determination of correction factors of 3H-β-self-absorption for quantitative evaluation of grain number in autoradiographic studies. Histochemistry 83, 65–70 (1985). https://doi.org/10.1007/BF00495302
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF00495302