User:LLLui/Cytochemistry/Bibliography

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The branch of histochemistry associated with the localization of cellular components by specific staining methods, now done mostly with fluorescently tagged monoclonal antibodies (immunocytochemistry).

Freeze fracture enzyme cytochemistry
Freeze-fracture cytochemistry (i.e., label-fracture and fracture-label), as developed by Pinto da Silva and co-workers (see a review"), has been a powerful technique that allows the cytochemical characterization of freeze-fractured membranes for the study of biological membrane ultrastructure in cell biology. In freeze-fracture cytochemistry, immunocytochemistry plays an important role for the visualization of cell membrane molecules by labeling with colloidal gold particles. Enzyme cytochemistry has received no attention although this can label many enzyme molecules in cells and tissues as well as immunocytochemistry can do. In this study, we show that enzyme cytochemistry can be applied to freeze-fracture cytochemistry.

In conclusion, we show new freeze-fracture cytochemistry based upon enzyme cytochemistry. This technique can identiiy and visualize the freeze-fractured membranes by labeling with an enzyme cytochemical marker (i.e. cerium phosphate reaction product). In addition, it may be possible to combine enzyme cytochemistry and fracture-flip for direct identification of enzyme molecules exposed on cell membranes. This technique should be a useful addition for analyzing the ultrastructure of various cell membranes.

This technique has, however, provided little information of the lipid protein compositions of cellular membranes. the combination of freeze-fracture replication with immunocytochemistry, is a sophisticated technique that can cytochemically define constituents of freeze-fractured membranes so that researchers can gain a better understanding of the distribution of the many molecules in biological membranes and identify the mem- branes of cell organelles

Origins
Cytochemistry, as defined by the present author, originated in the period 1930-1940 from a number of centres. It owes much to the earlier work of Brachet in Brussels (as reviewed by Brachet 1950), who used a mixture of methyl green and pyronin to demonstrate the localization, and relative abundance, of DNA and RNA in cells. He controlled the use of these dyes by the action of specific nucleases (e. g. Brachet 1954; also Jacobson and Webb 1952). This work demonstrated that there was not a specifically animal nucleic acid (thymonucleic acid) and a specifically plant nucleic acid (phytonucleic acid) as had been thought by biochemists who had found that the animal tissue most rich in nucleic acid, namely the thymus, contained what we now call deoxyribonucleic acid (DNA) whereas the richest plant source of nucleic acid, namely yeast, contained ribonucleic acid (RNA). Brachet (1950) showed that both occurred in nucleated cells, whether of plants or animals.

Much of the spirit of cytochemistry, particularly the need for highly sensitive, very precise, chemical methods which would be sufficiently sensitive to allow biochemical activity to be related to detailed histology, came from Linderstrom-Lang and Holter in the Carlsberg Laboratory in Copenhagen (see Holter and Moller 1976). This laboratory emphasized the relationship between structure and function and one of the most important concepts of cellular biology, namely endocytosis, came from this group (e. g. Holter 1961, 1965). A practical outcome oftheir studies was the development of a cryostat microtome (Linderstrom-Lang and Mogensen 1938) which is now an essential tool in cytochemistry.

Aims
The aims and outlook of cytochemistry are to study the biochemistry of individual cells within a tissue which may contain several different cell types. It is an extension of conventional biochemistry. But it has a further facet, namely that it is a nondestructive method of analysis. Thus whereas Glick's quantitative histochemistry may involve the destruction of the structure of the tissue, as does homogenate biochemistry, cytochemistry tries to retain the structure intact and so to test the biochemical activity of intact cellular systems rather than the biochemical behaviour of these systems after homogenization, and isolation into a foreign medium, have distorted membrane structures and spatial relationships. Thus its outlook is almost the reverse of that of histochemistry, as defined by Barka and Anderson (1963).

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