Practical Histochemitry

Part 1 The preparation of sections: freeze-sectioning procedure - methods, to chill tissue, to cut sections, to pick up the sections, to mount the tissue, general advice on sectioning. Part 2 Methods of incubating sections: methods of incubation; the need for stabilizers; the use of tissue stabilizers; method of assessing the efficacy of a colloid-stabilizer; the selection of the colloid stabilizer; cytochemistry of isolated cells; ions of stabilizing sections; to prepare a stock solution of polyvinyl alcohol. Part 3 Methods of quantification: elution techniques; elution method; precise quantification-scanning and integrating microdensitometry; assay by microdensitometry; polarized light microscopy; types of birefringence; characteristic features in polarized light microscopy; applications of polarized light microscopy; induce birefringence. Part 4 Common histological stains: special methods of preparing tissue; fixatives; staining procedures. Part 5 The analysis of chemical components of cells and tissues: general analysis - the determination of the major classes of compound present, proteins, acidic substances, carbohydrates, lipids, minor components; practical methods - reactions for protein, reactions for nucelic acids and polyphosphate, reactions for polysaccharides, reactions for lipids, minor components. Part 6 Enzyme histochemistry: characteristics of the histochemical reactions for enzymatic activity; validity of histochemical enzymatic reactions; studies on bound or protected enzymes; the question of the use of stabilizers for assaying bound enzymes studies on intact cells; classification of enzymes - phosphatases, aminopeptidases and napthylamidases, B glucoronidase, carbonic anhydrase, n-acetyl - B-glucosaminidases, oxidases, peroxidases, succinate dehydrogenase, lactate dehydrogenases, glutamate dehydrogenases, B-hydroxyacyl CoA dehydrogenase, steroid dehydrogenases, NADP-dependent dehydrogenases, NADP-diaphorase. Appendices: effect of fixation on enzymes; buffers.