The histochemical basis of quantitative histology*

Abstract

KEY W 0 RD S . Enzymes, cell function, histochemistry, histology, image analysis, muscle. SUMMARYThe interaction between histochemistry and microscopy for quantifying the morphology and function of cells in sections of tissue is reviewed.In principle, the morphological parameters of cells can be measured in suitably-stained sections using image analysers. In practice, however, the individual cells often cannot be distinguished from each other in such systems. This problem might be overcome, at least in muscle, by visualizing types I11 and IV collagen in the connective tissue and basement membranes surrounding the cells.The functional capacities of cells within tissues can be inferred by measuring the activities of their key enzymes with quantitative histochemical techniques. At present, the wrong enzymes are frequently chosen for this purpose. Flux-generating and physiologically significant enzymes are preferable, but this requires new histochemical techniques to be devised and validated quantitatively for use with the computer-assisted analytical microscopes that are now becoming available commercially.

Digital imaging fluorescence microscopy: spatial heterogeneity of photobleaching rate constants in individual cells.

“…This strategy provides an objective means to compare video equipment and to maximize control of dayto-day variables such as lens alignment, lamp stability, and focus. Moreover, it furnishes a means to initiate the development of radiometric and geometric microscopic fluorescence standards and, finally, allows the application of a whole range of image processing and image analysis methodology (22)(23)(24)(25)(26)(46)(47)(48)(49)(50).…”

“…In general, these measurements have been restricted to and integrated over a small area of the microscope field, with little effort to retain spatial information. A number of recent efforts that use low light level cameras and video digital image processing devices have been directed at obtaining quantitative information on the spatial, spectral, and radiometric distribution of fluorophores within single cells (14,17,(22)(23)(24)(25)(26)(27)(28). Benson et al (14) have demonstrated that an image processor incorporated into a video-intensified microscopy system can provide a quantitative measure of the spatial distribution of the fluorescence intensity and can be used to determine oxygen concentration by fluorescence quenching.…”

Adaptive sex-dependent changes in the zonation of carbohydrate and lipid metabolism in rat liver lobules after partial hepatectomy

“…Anesthesia was induced with ether (approximately 2 min) followed by sodium pentobarbital (Nembutal; 40 mg/kg body weight, intraperitoneally). At 0, 4,8,24,48,336 and 720 hr after partial hepatectomy, the livers were removed immediately and small pieces up to 5 mm3 were chilled in isopentane cooled with liquid nitrogen and stored at -80" C until further use. Animal welfare was in accordance with the guidelines of the University of Amsterdam.…”

Age- and Strain-Related Differences in Dehydrogenase Activity and Glycogen Levels in CBA and C57 Mouse Cochleas

“…For instance, when succinate, one of the intermediate products in the citric acid cycle, is included in the incubation medium, SDH will catalyze its oxidation to fumarate [Clarke et al, 1989;Spector, 1975;Vosteen, 1960] and the liberated hydrogen ion will reduce the electron-accepting tetrazolium salt to an insoluble, blue formazan [Koide et al, 1964;Spoendlin and Balogh, 1963;Vosteen, 1960;Yang et al, 1990]. Thus, the density of the reaction product formed is proportional to a cell's oxygen consumption and energy production and can be used to infer aerobic metabolic activity [Chalmers et al, 1992;Peter et al, 1972;Spoendlin and Balogh, 1963;Stoward and Ploem, 1982;Vosteen, 1960;Yang and Han, 1991;Yang et al, 1990].…”