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Principle and method of kinetic microphotometric enzyme activity determination in situ

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Summary

An advanced apparative set-up is described for multipositional microphotometric recording of histochemical enzyme reactions in cryostat sections. It consists of a computer controlled microscope photometer with scanning stage. Measurements on the same tissue section may be performed at 12 preselected positions. These are repeatedly brought into the measuring beam in several measuring cycles. The complete measuring process, storage of measuring position coordinates, movements of the stage and statistical evaluation of the data is under computer control. By use of the gel film technique, extinction changes in tetrazolium coupled enzyme reactions can be measured continuously at initial rate conditions. Measurements are performed at identical conditions and can thus be analysed as relative enzyme activities.

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References

  • Altman, F.P.: The use of a recording microdensitometer for the quantitative measurement of enzyme activities inside tissue sections. Histochemie 27, 125–136 (1971)

    Google Scholar 

  • Altman, F.P.: Quantitative microscopy of enzyme reactions in tissue sections. Microsc. Acta 79, 327–338 (1977)

    Google Scholar 

  • Butcher, R.G.: Precise cytochemical measurement of neotetrazolium formazan by scanning and integrating microdensitometry. Histochemie 32, 171–190 (1972)

    Google Scholar 

  • Butcher, R.G.: The measurement in tissue section of the two formazans derived from Nitroblue tetrazolium in dehydrogenase reactions. Histochem. J. 10, 739–744 (1978)

    Google Scholar 

  • Butcher, R.G., Altman, F.P.: Studies on the reduction of tetrazolium salts. II. The measurement of the half-reduced and fully reduced formazans of neotetrazolium chloride in tissue sections. Histochemie 37, 351–363 (1973)

    Google Scholar 

  • Brumberg, V.A., Pevzner, L.Z.: Cytospectrophotometric studies on the lactate dehydrogenase isozymes in functionally different neuron-neuroglia units. I. Optimal procedure conditions and topochemical comparisons in normal mice. Acta Histochem. (Jena) 54, 218–229 (1975)

    Google Scholar 

  • Bykov, V.L., Prianishnikov, V.A.: Quantitative histochemical and stereological study of alkaline phosphatase in the rat thyroid gland. Histochemistry 58, 129–134 (1978)

    Google Scholar 

  • Felgenheuer, K., Glenner, G.G.: Quantitation of tissue bound renal aminopeptidase by a microdensitometric technique. J. Histochem. Cytochem. 14, 53–63 (1966)

    Google Scholar 

  • Frasch, A.C.C., Itoiz, M.E., Cabrini, R.L.: Microspectrophotometric quantitation of the diaminobenzidine reaction for histochemical demonstration of cytochrome oxidase activity. J. Histochem. Cytochem. 26, 157–162 (1978)

    Google Scholar 

  • Galustyan, G.E., Prianishnikov, V.A.: Cytospectrophotometric study of the kinetics of histochemical reaction of GABA-transaminase in cryostat sections of rat cerebellar cortex. Histochemistry 57, 77–86 (1978)

    Google Scholar 

  • Garcia, A.M.: Studies on DNA in leucocytes and related cells of mammals. II. On the Feulgen reaction and two-wave-length microspectrometry. Histochemie 3, 178–196 (1962)

    Google Scholar 

  • Goldspink, G.: Succinic dehydrogenase content of individual muscle fibres at different ages and stages of growth. Life Sci. 8, 791–808 (1969)

    Google Scholar 

  • Howells, K.F., Branford White, C.J., Goldspink, G.: Zytophotometrische Bestimmungen des Bernsteinsäure-Dehydrogenase-Gehalts von Muskelfasern mit dem LEITZ-Mikroskop-Photometer MPV. LEITZ-Mitt. Wiss. Tech. 7, 77–79 (1978)

    Google Scholar 

  • Mayall, B.H., Mendelsohn, M.L.: Errors in absorption cytophotometry: Some theoretical and practical considerations. In: Introduction to quantitative cytochemistry-II., Wied, G.L., Bahr, G.F. (eds.), pp. 171–197, New York, London: Academic Press 1970

    Google Scholar 

  • Nolte, J., Pette, D.: Quantitative microscope-photometric determinations of enzyme activities in cryostat sections. In: Recent advances in quantitative histo- and cytochemistry. Dubach, J., Schmidt, U. (eds.), pp. 54–60. Bern, Stuttgart, Wien: Hans Huber 1971

    Google Scholar 

  • Nolte, J., Pette, D.: Microphometric determination of enzyme activity in single cells in cryostat sections. I. Application of the gel film technique to microphotometry and studies on the intralobular distribution of succinate dehydrogenase and lactate dehydrogenase activities in rat liver. J. Histochem. Cytochem. 20, 567–576 (1972a)

    Google Scholar 

  • Nolte, J., Pette, D.: Microphotometric determination of enzyme activity in single cells in cryostat sections. II. Succinate dehydrogenase, lactate dehydrogenase and triosephosphate dehydrogenase activities in red, intermediate and white fibres of soleus and rectus femoris muscles of rat. J. Histochem. Cytochem. 20, 577–582 (1972b)

    Google Scholar 

  • Pearse, A.G.E.: “Histochemistry, Theoretical and Applied”, 3rd edition. Edinburgh and London: Churchill Livingstone 1972

    Google Scholar 

  • Pette, D.: Semiquantitative microphotometric enzyme activity determination by means of comparative and multipositional recording of reaction kinetics, p. 274. Proceedings Vth International Congress Histochemistry and Cytochemistry, Bucharest, 1976

  • Pette, D., Brandau, H.: Intracellular localization of glycolytic enzymes in cross-striated muscles of locusta migratoria. Biochem. Biophys. Res. Commun. 9, 367–370 (1962)

    Google Scholar 

  • Pette, D., Smith, M.E., Staudte, H.W., Vrbová, G.: Effects of long-term electrical stimulation on some contractile and metabolic characteristics of fast rabbit muscles. Pflügers Arch. 338, 257–272 (1973)

    Google Scholar 

  • Pette, D., Wimmer, M.: Kinetic microphotometric activity determination in enzyme containing gels and model studies with tissue sections. Histochemistry 64, 11–22 (1979)

    Google Scholar 

  • van der Ploeg, M., van Duijn, P.: Cytophotometric determinations of alkaline phosphatase activity of individual neutrophilic leucocytes with a biochemically calibrated model system. J. Histochem. Cytochem. 16, 693–706 (1968)

    Google Scholar 

  • Rosenquist, T.H.: On the use of quantitations in histochemistry: a review of the literature. Histochem. J. 8, 205–208 (1976)

    Google Scholar 

  • Täljedal, I.B.: Direct fluorophotometric recording of enzyme kinetics in cryostat sections. Glucose 6-phosphate dehydrogenase in the endocrine mouse pancreas. Histochemie 21, 307–313 (1970)

    Google Scholar 

  • Wenk, H., Krug, H.: Zur histophotometrischen Quantifizierung von Enzymreaktionen, dargestellt am Beispiel von Acetylcholinesterase. Acta Histochem. (Jena) Suppl. 15, 433–440 (1975)

    Google Scholar 

  • Wimmer, M., Pette, D.: Microphotometric studies on intraacinar enzyme distribution in rat liver. Histochemistry 64, 23–33 (1979)

    Google Scholar 

  • Zimmermann, P.: Cytophotometric studies on the reaction kinetics of acid phosphatase in motoneurons marked by injection of Evans blue into the soleus muscle of the mouse. Cell Tissue Res. 178, 239–247 (1977)

    Google Scholar 

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Pette, D., Wasmund, H. & Wimmer, M. Principle and method of kinetic microphotometric enzyme activity determination in situ. Histochemistry 64, 1–10 (1979). https://doi.org/10.1007/BF00493350

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