Practical Histochemitry

Many applied biological and medical scientists have asked how they could apply histochemistry to their particular studies. Often their questions were purely technical - how to stain for a particular enzyme or substance. They saw histochemistry as an extension of hisology and they wanted a well established recipe. They did not want to go into the science underlying the staining method any more than they did when applying special stains in histology. This book aims to give them a technique that they can follow as simply as a routine histological method which has a scientific basis. Other workers had problems in tissue metabolism and chemical dysfunction. For these, histochemistry was an extension of biochemistry. Its special advantages were that it allowed them to relate a specific activity to a particular tissue component and it was a form of biochemistry that could be done on relatively minute pieces of tissue such as could be removed safely by biopsy, or could be kept either in maintenance or proliferative culture. But they too did not want to delve into the variants that can be found for almost every histochemical reaction. This book is a collection of histochemical reactions with simple techniques which, if followed precisely, will yield reactions where the reactive compounds or enzymes are present, and will give a measure of the degree of activity actually shown in the tissue at the time of its removal, and can also indicate the total activity of which the tissue could be capable.

Table of Contents:
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.