Summary
The effect of storage of unfixed cryostat sections from rat liver for 4 h, 24 h, 3 days and 7 days at -25°C was studied on the activities of lactate dehydrogenase, glucose-6-phosphate dehydrogenase, xanthine oxidoreductase, glutamate dehydrogenase, succinate dehydrogenase (all demonstrated with tetrazolium salt procedures), glucose-6-phosphatase (cerium-diaminobenzidine method), 5′-nucleotidase (lead salt method), dipeptidyl peptidase II, acid phosphatase (both simultaneous azo coupling methods), d-amino acid oxidase (cerium-diaminobenzidine-cobalt-hydrogen peroxide procedure) and catalase (diaminobenzidine method). The effect of drying of the cryostat sections at room temperature for 5 and 60 min was investigated as well. The enzyme activities were quantified by cytophotometric measurements of test and control reactions. The test minus control reaction was taken as a measure for specific enzyme activity. It was found that the activities of all the enzymes investigated, with one exception, were affected neither by storage of the cryostat sections at -25°C for up to 7 days, nor by drying of the sections at room temperature for up to 60 min. The exception was xanthine oxidoreductase, whose activity was reduced by 20% after 5 min drying of sections or after 4 h storage. Therefore, only incubations for xanthine oxidoreductase activity have to be performed immediately after cutting cryostat sections, whereas for the other enzymes a considerable margin appears to exist.
Similar content being viewed by others
References
Frederiks, W. M. & Marx, F. (1987) Changes in cytoplasmic and mitochondrial enzymes in rat liver after ischemia followed by reperfusion. Exp. Mol. Pathol. 47, 291–9.
Frederiks, W. M. & Marx, F. (1988) A quantitative histochemical study of 5′-nucleotidase activity in rat liver using the lead salt method and polyvinyl alcohol. Histochem. J. 20, 207–14.
Frederiks, W. M. & Marx, F. (1989) Changes in acid phosphatase activity in rat liver after ischemia. Histochemistry 93, 161–6.
Frederiks, W. M., Myagkaya, G. L., Fronik, G. M., VanVeen, H. A., Vogels, I. M. C. & James, J. (1983) The value of enzyme leakage for the prediction of necrosis in liver ischemia. Histochemistry 78, 459–72.
Frederiks, W. M., Marx, F. & Myagkaya, G. L. (1986) A histochemical study of changes in mitochondrial enzyme activities of rat liver after ischemia in vitro. Virchows Arch. (Cell Pathol.) 51, 321–9.
Gossrau, R. & Lojda, Z. (1980) Study on dipeptidylpeptidase II (DPP II). Histochemistry 70, 53–76.
Henderson, B., Loveridge, N., Robertson, W. R. & Smith, M. T. (1981) The influence on enzyme activity of storage of tissue blocks at -70°C. A quantitative cytochemical study. Histochemistry 72, 545–50.
Jonges, G. N., VanNoorden, C. J. F. & Gossrau, R. (1990) Quantitative histochemical analysis of glucose-6-phosphatase activity in rat liver using an optimized ceriumdiaminobenzidine method. J. Histochem. Cytochem. 34, 1413–9.
Kooij, A., Frederiks, W. M., Gossrau, R. & VanNoorden, C. J. F. (1991) Localization of xanthine oxidoreductase activity using the tissue protectant polyvinyl alcohol and final electron acceptor tetranitro BT. J. Histochem. Cytochem. 39, 87–93.
Patel, H. R. H., Frederiks, W. M., Marx, F., Best, A. J. & VanNoorden, C. J. F. (1991) A quantitative histochemical study of d-amino acid oxidase activity in rat liver in relationship with feeding conditions. J. Histochem. Cytochem. 39, 81–6.
Roels, F., Wisse, E., De Prest, B. & Van DerMeulen, J. (1975) Cytochemical discrimination between catalases and peroxidases using diamenobenzidine. Histochemistry 41, 281–312.
Stirpe, F. & Della Corte, E. (1969) The regulation of rat liver xanthine oxidase: conversion in vitro of the enzyme activity from dehydrogenase (type D) to oxidase (type O). J. Biol. Chem. 244, 3855–63.
VanNoorden, C. J. F. (1984) Histochemistry and cytochemistry of glucose-6-phosphate dehydrogenase. Progr. Histochem. Cytochem. 15.4, 1–85.
VanNoorden, C. J. F. & Butcher, R. G. (1991) Quantitative enzyme histochemistry. In Histochemistry: Theoretical and Applied edited by Stoward, P. J. & Pearse, A. G. E., Vol 3, 4th edn pp 355–432. Edinburgh: Churchill Livingstone.
VanNoorden, C. J. F. & Frederiks, W. M. (1992) Enzyme Histochemistry: a Laboratory Manual of Current Methods. Oxford: Oxford University Press.
Wachstein, M. & Meisel, E. Histochemistry of hepatic phosphatases at a physiological pH. Am. J. Clin. Path. 27, 13–23.
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Frederiks, W.M., Ouwerkerk, I.J.M., Bosch, K.S. et al. The effects of storage on the retention of enzyme activity in cryostat sections. A quantitative histochemical study on rat liver. Histochem J 25, 119–122 (1993). https://doi.org/10.1007/BF00157983
Received:
Revised:
Issue Date:
DOI: https://doi.org/10.1007/BF00157983