Histological Techniques, Softcover reprint of the original 1st ed. 1976

'Eyw rpwvry ~OiWTO~ EV Tli E(}'rllup JOHN, J, 23 A SURVEY OF THE PRINCIPAL WORKS on histological technique that have th appeared since the end of the 19 century shows such a diversity of concept and directive ideas, in spite of the fundamental unity of subject, that the choice of the underlying orientation was perhaps the most difficult problem facing me when MM. Masson & Cie, requested me to write this book. A classification of these works based on their particular orientation and carrying the excess inherent in all that is schematic would lead to the definition of three types of books. Some are handbooks or treatises on "microscopy". They are tacitly limited to the biological applications of the microscope, all of which are reviewed, and include in the same volume the techniques of histology proper along with those of bacteriology, botany and embryology. Such a work, on the scale of an encyclopaedic treatise contributed by specialists from a number of discip­ lines and meant to be consulted as a dictionary, may have a certain value if it is really exhaustive and periodically revised. But it is no longer possible on the scale of a handbook. A single person, however gifted, cannot have acquired sufficient mastery over all the techniques of the above disciplines to be able to speak from personal experience.

Foreword.- Note concerning textual conventions.- Histological research and diagnostic histology.- Automatic, easy, difficult and capricious technique.- Choice of a technique.- Conclusion of technique.- one General Principles of Histological Technique.- 1. — Vital examination.- Removal of pieces.- Making preparations.- Examination under the light microscope.- Examination under the dark-field microscope.- Examination under the phase-contrast and interference microscope.- 2. — Vital Staining.- Definition.- Objects, advantages and disadvantages.- Classification of vital stains.- Theories of vital staining.- General rules for applying vital stains.- 3. — Fixation.- Classification and mode of action of fixatives.- Fixation by physical agents.- Fixation by chemical agents.- General remarks on fixative mixtures.- Fixative mixtures containing an “indifferent” salt.- Mixtures containing several fixing agents.- Period of fixation.- Temperature of fixation.- Modes of fixation.- Fixation with vapours.- Fixation in liquids.- Choice of fixative.- 4. — Embedding and preparatory operations.- Termination of fixation.- Embedding in paraffin.- Properties of the embedding medium.- Procedure of paraffin embedding.- Choice of method for paraffin embedding and summary of operating procedure.- Embedding in nitrocellulose (celloidin, collodion).- Properties of the embedding mass.- Procedure of embedding.- Embedding in gelatin.- Embedding in gelatin after Apathy.- Embedding in gelatin after Heringa and Ten Berge.- Embedding in gelatin after Baker.- Embedding in gelatin after Pearse.- Other methods of embedding.- The solid polyethylene-glycols.- Steedman’s ester-wax.- Double embedding.- Embedding in agar-paraffin.- Embedding in celloidin-paraffin: the method of Apathy—the method of Pfuhl.- 5. — Cutting and sticking sections.- The razor.- The razor-holder.- The object-holder.- The mechanism of advance.- Choice of a microtome.- Maintenance of the microtome.- Cutting and handling paraffin sections.- Preparing the block.- Cutting sections.- Handling and spreading sections.- Cutting sections in paraffin-agar.- Cutting and handling celloidin and celloidin-paraffin sections.- Preparation of celloidin blocks to be cut under alcohol.- Preparation of celloiding blocks to be cut under terpineol.- Preparation of celloidin-paraffin blocks.- Cutting sections.- Sticking sections.- Cutting and handling gelatin sections.- Cutting and handling frozen sections.- Preparing blocks.- Cutting sections.- Handling and sticking sections.- 6. — Staining and mounting microscopical preparations.- Definition and nomenclature.- General theory of staining and the classification of stains.- Theories of histological staining.- The chemical theory.- The concept of staining by inhibition and by precipitation.- Adsorption as a factor in the binding of stains.- Electrostatic adsorption as a mechanism of histological staining.- Theory of indirect staining.- The density of structures as a factor in histological staining.- The texture of tissue constituents and their penetration by stains.- General remarks on the practice of staining.- Choice of stains.- Preparation of solutions.- Glassware.- Deparaffining sections.- Collodioning sections.- Dissolving crystals of mercury.- Summary of operations preceding the staining of sections.- Mounting media miscible in water: Apathy’s syrup; The glycerine gum; Laevulose syrup; Gelatinized glycerine.- Mounting media miscible with benzenic hydrocarbons.- Summary of different mounting procedures.- Luting the preparations.- Conservation of microscopical preparations.- 7. — The examination of histological preparations.- The graphical representation of histological preparations.- The reconstruction of histological preparations.- Graphical reconstruction.- Plastic reconstruction.- The quantitative study of histological preparations.- The causes of error in measuring under the microscope.- Measuring lengths in the plane of the stage.- Measuring lengths along the optical axis of the microscope.- Measuring areas perpendicular to the optical axis of the microscope.- Procedures of linear integration to measure areas and relative volumes.- Appendix. — List of principal dye-stuffs used in histology.- Anthraquinone.- Azins.- Azo-dyes.- Fluorochromes— Indulins—Natural dyes.- Nitro-dyes—Nitroso-dyes—Phenyl methanes.- Phthalocyanin— Pyrazolones—Quinolines—Thiazins.- Xanthenes.- two General Methods.- 8. — Topographical fixation.- The principal topographical fixatives.- Aqueous liquids.- Alcoholic liquids.- The practice of topographical fixation.- Choice of fixative.- Removal and fixation.- Stopping fixation and storing pieces.- Choice of embedding method.- 9. — Topographical staining.- Classification of topographical stains.- Nuclear staining by progressive haematoxylin lakes.- Aluminium lakes ofhaematein.- Progressive ferric lakes ofhaematein.- Progressive chromic lakes ofhaematein.- Selective staining of collagen fibres.- Staining based on competition between two acidic dye-stuffs.- Haemalum-picro-indigocarmine.- Ramon y Cajal’s trichrome.- Nuclear fast-red picro-indigocarmine.- Van Giesons’ method.- Variants of Van Gieson’s method.- The method of Curtis.- Variants of the method of Curtis.- Gaussen’s histo-polychrome.- A. Prenant’s triple stain.- Advantages and disadvantages of methods based on competition between two acidic stains.- Staining that involves phosphomolybdic or phosphotungstic acid.- Original method of Mallory.- Masson’s trichrome.- Gomori’s trichrome.- Variants of Gomori’s trichrome.- One-step trichrome: Gabe and Ms Martoja.- Heidenhain’s azan.- Variants of the azan stain.- Petersen’s method.- Advantages and disadvantages of methods involving phosphomolybdic or phosphotungstic acids.- Staining based on the demonstration of cytoplasmic tinctorial affinities.- Nuclear staining with a haematoxylin lake followed by counter-staining with an acid dyestuff.- Millot’s triple staining.- Safranin-light green.- The Ehrlich-Biondi-Heidenhain method.- Mann’s technique.- Dobell’s variant.- The Mann-Dominici method.- Staining with azur eosinates.- Advantages and disadvantages of the methods of this group.- Bulk staining.- Coloration with the aluminium lake of gallamine blue.- Staining with boracic carmine.- Staining with Carmalum.- Bulk staining with P. Mayer’s haemalum.- Staining with haematein IA of Apathy.- 10. — Decalcifying and softening very hard tissues.- Decalcifying.- Nitric acid—Sulfurous acid—Trichloracetic acid.- Formic acid— Chromic acid —Picric acid —Sodium ethylene-diamino-tetràcetate (EDTA, versene)—Decalcifying by organic buffers.- Softening cuticular structures and keratin.- Chlorine dioxide.- Murrays’ method.- three Histochemical Methods.- 11. — General introduction to the study of histochemical techniques.- Definitions.- Current progress in histochemical research and the ways in which its techniques may be employed.- The Requirements Exacted by Histochemical Investigations.- Chemical conditions.- Morphological conditions.- 12. — Histophysical methods.- Absorption spectrophotometry and histophotometry.- Emission histospectrography.- Fluoroscopy and fluorescence spectrography.- Apparatus.- Fluoroscopy as a morphological technique.- Primary fluorescence for histochemical identification.- Secondary fluorescence following histochemical reactions.- Immuno-histochemistry.- Historadiography.- Phase contrast microscopy and interference microscopy.- Microscopy in polarised light.- Autoradiography.- 13. — Histochemical detection of mineral substances.- Micro-incineration.- Fixation—Preparation and spreading of sections.- Conduct of incineration.- Mounting and examination of spodograms.- Identification of mineral matter in spodograms.- Detection of mineral anions.- Chlorides.- Iodides and Thyroid iodine.- Phosphorus.- Sulphur.- Arsenic.- Silicon.- Detection of mineral cations.- Sodium and Potassium.- Calcium.- Barium and strontium.- Magnesium.- Zinc.- Aluminium.- Iron.- Nickel and cobalt.- Copper.- Mercury.- Silver.- Gold.- Bismuth and Lead.- 14. — Histochemical detection of properties common to several radicals and functional groups.- Histochemical detection of reducing compounds.- The ferric ferricyanide reaction:.- The reduction of silver salts.- The reduction of tetrazolium salts.- The histochemical study of acidophilia.- The histochemical study of basophilia.- The physico-chemical factors influencing basophilia:.- The detection of basophilia for histochemical purposes:.- Estimation of the tissue zone containing proteins at isoelectric point.- The histochemical interpretation of basophilia in tissue.- The histochemical study of metachromasia.- Metachromatic stains.- The mechanism of metachromasia.- The practical conduct of the metachromatic reaction.- Histochemical interpretation of the metachromatic reaction.- Other staining indicative of the presence of anions:.- Paraldehyde fuchsin.- Paraldehyde thionine.- Phthalocyanins (alcian blue, alcian green, alcian yellow, astra blue).- 15. — Histochemical detection of aldehydes and ketones.- The Schiff reagent.- The preparation of Schiffs reagent.- Alkaline silver complexes.- Benzidine and O-dianisidine.- Phenylhydrazine.- 2-hydroxy-3-naphthoic acid hydrazide.- p-Phenylenediamine.- Blocking reactions of the aldehyde groups:.- Aniline chlorhydrate. Hydroxylamine. Semicarbazide. Thiosemicarba- zide. Phenylhydrazine. Sodium bisulphite.- The distinction between aldehydes and ketones.- 16. — Histochemical detection of phenols and naphthols.- The azo-reaction.- The reagents.- Lison’s technique for the preparation of diazonium salts.- Technique of Lillie and al. for the preparation of the diazo salt of safranine 0.- Technique of Lillie and Glenner for the preparation of the diazo salts of S acid (8-amino-naphthol-5 sulphonic acid).- Davis’s technique for the diazotisation of pararosaniline.- Coupling.- Histochemical interpretation.- The Indo-reaction.- The Gibbs reaction.- The argentaffin reaction.- The phaeochrome (chromaffin) reaction.- 17. — Histochemical detection of carbohydrates.- Histochemical detection of glucose.- Histochemical detection of ascorbic acid.- Demonstration of ascorbic acid: in pieces of tissue, after freeze drying.- and on cryostat section.- Histochemical Detection of Polysaccharides.- Classification of polysaccharides detectable by histochemical methods.- Techniques for the detection of polysaccharides.- Oxidative reactions:.- Reagents and working technique.- Metachromatic reactions.- Sulfuric and phosphoric esterification.- Fixation of metallic ions.- Combination of the colloidal iron technique with the PAS-reaction.- Marker stainings.- Histochemical identification of different kinds of polysaccharides.- The practice of histochemical tests for polysaccharides.- 18. — Histochemical detection of fats and related compounds.- Classification of fats.- Preparation of the tissues for a histochemical study of lipids.- The histochemical detection of lipids by means of lysochromes.- By lysochromes in ethanol solution.- by lysochromes in supersaturated isopropyl alcohol solution.- by fluorescent lysochromes.- The use of extraction techniques in the histochemical study of lipids.- Ciaccio’s technique for selectively rendering insoluble the phospho-and sphingolipids.- Keilig’s technique for the differential extraction of lipids.- Histochemical reactions and stains permitting the identification of certain lipids.- Optical anisotropy.- Demonstration of the acid nature of lipids.- Demonstration of the unsaturated nature of lipids.- Demonstration of the complexes formed by lipids with chromium compounds.- Demonstration of lipids bearing carbonyl groups.- Demonstration of carbohydrate constituents of certain lipids.- Demonstration of the sterol radical.- Histochemical Characteristics of the Principal Categories of Lipids.- Glycerides.- Phosphatide esters.- Plasmalogens.- Sphin-golipids.- Steroids.- Carotenoids.- Chromolipoids.- Ceroid pigment.- Lipofuscins.- Haemofuscin.- Dichotomous Keys for the Analysis of Lipids.- 19. — The histochemical detection of amino-acids and proteins.- The Fixation of Proteins.- The Histochemical Reactions of Proteins as Such.- The Histochemical Detection of the A-Amino-Acid Group.- Reactions depending on the formation of azomethines.- Reactions of condensation with dinitrofluorobenzene.- Reactions depending on oxidative deamination.- Blocking reactions for primary amine groups.- Reactions for the selective detection of certain amino-acids.- Demonstration of the guanidyl radical and of arginine.- Demonstration of the imidazol group.- Demonstration of tyrosine.- Demonstration of indole groups and in particular of tryptophane.- Demonstration of thiol and disulphide groups.- Demonstration of electropolar features of proteins.- Tests for acidophilia as a method for the identification of proteins.- The evaluation of the zone of the isoelectric point in the histochemical study of proteins.- The identification of certain proteins by enzymatic digestion.- Trypsine, chrymotrypsine, pepsine.- Collagenase.- Elastase.- The identification of proteins by differential solubility.- 20. — Histochemkal detection of nucleoproteins and metalloproteins.- Histochemical Detection of Nucleoproteins.- Demonstration of nucleic acids through their ultraviolet absorption.- Demonstration of nucleic acids through their basophilia.- Demonstration of nucleic acids by staining with two basic dyes.- Nucleal reactions.- Techniques for extraction of nucleic acids.- Histochemical Detection of Metalloproteins.- Histospectroscopy.- Pseudo-peroxidase properties.- 21. — Histochemkal detection of some products of protein metabolism.- Compounds associated with the metabolism of phenylalanine and tyrosine.- Fluoroscopic detection of biogenic monoamines.- Histochemkal detection of catecholamines.- Histochemkal detection ofmelanins.- Methods for bleaching.- Compounds related to the metabolism of tryptophane.- Compounds related to the metabolism of histidine.- Compounds related to the metabolism of haemoglobin.- Hemosiderins.- Bile pigments.- Porphyrins.- Waste-products of protein metabolism.- The histochemkal detection of urea.- Histochemkal detection of purines.- 22. — The histochemkal detection of the principal enzymes.- The preparation of tissues intended for histo-enzymological research.- The general course of histo-enzymological reactions.- Histochemical Detection of Hydrolases.- Phosphatases.- 5-Nucleotidase (Adenosine 5-phosphatase).- Gomori’s technique.- Wachstein and Meisel technique.- Non-specific acid Phosphomonoesterase.- Gomori’s technique using lead nitrate.- Techniques using naphthol phosphates.- Phosphamidase.- Technique of Gomori.- Meyer and Weinnjann’s technique.- Glucose-6-Phosphatase.- Chiquoine’s technique.- Wachstein and Meisel’s technique.- Carboxylic esterases.- Cholinesterases.- Coer’s technique.- Arvy’s technique.- The technique of Wachstein and al..- Non-specific esterases.- Preparation of tissues.- Techniques involving naphthylacetates.- Techniques involving indoxyl acetates.- Lipases.- Sulphatases.- Glucosidases (glycosidases).- Carbonic anhydrase.- Peptidases.- Deoxyribonuclease.- Histochemical Detection of Transferases.- Histochemical Detection of Oxido-Reductases.- Oxidases.- Peroxidases.- Aerobic dehydrogenases.- Anaerobic dehydrogenases.- Four The Methods of General Cytology.- 23. — Introduction to the techniques of general cytology.- The problem of cytological fixation.- Chromo-osmic solutions.- Chromic fluids without osmium tetroxide and acetic acid.- Fluids based on heavy metals other than chromium and osmium.- Chromo-acetic fluids.- Regressive staining with haematoxylin lakes.- Ferric lakes.- Heidenhain’s technique and its variants.- Dobell-Hirschler haematoxylin.- Cupric lakes.- 24. — Techniques for studying the nucleus.- The general study of nuclear structures.- Vital observation.- Vital stains.- Nuclear structures in smears.- Nuclear structures on sections.- 25. — Techniques for demonstrating the golgi apparatus.- Osmic impregnation.- Silver impregnation.- Choice between osmic and silver impregnation.- Demonstration of the Golgi apparatus by staining or histochemical reactions.- 26. — Techniques for demonstrating the chondriome.- Mitochondrial fixations.- Preparation of pieces for mitochondrial staining.- Mitochondrial stains.- Staining with acidfuchsin.- Staining with crystal violet.- Regressive staining of the chondriosomes with the haematoxylin lakes.- Silver impregnation of the chondriome.- Choice of mitochondrial techniques.- 27 — Techniques for demonstrating the ergastoplasm and related structures.- Ergastoplasm and ergastoplasmic parasomes.- Techniques for demonstrating yolk nuclei.- 28. — Techniques for studying the centrosome and fibrillar differentiation of the cytoplasm.- Techniques for demonstrating the centrosome.- Techniques for demonstrating tonofibrils.- Techniques for brush borders and striated borders.- Techniques for demonstrating cilia.- 29. — Techniques for demonstrating secretory granules.- Topographical stains.- Cytological techniques.- Staining with neutral dyes.- GomorVs method and its variants.- Chromic haematoxylin phloxine.- The Mann-Dominici stain after permanganate oxidation.- the paraldehyde-fuchsin, Groat’s haematoxylin and picro-indigocarmine stain.- Method of Solcia, Vassallo and Capella:.- Staining with toluidine blue.- Staining with astra blue.- Staining with pseudo-isocyanin.- Silver impregnation techniques.- Distinction between secretory granules and lysosomes.- Five Histological Examination of the Principal Tissues and Organs.- 30. — Techniques for isolation and maceration and the demonstration of cell boundary.- Maceration techniques.- Demonstration of cell boundaries on spread membranes.- Demonstration of cell boundaries by bulk staining.- Demonstration of cell boundaries on paraffin wax sections.- 31. — Techniques for the histological study of the integument and of its outgrowths.- The integument of vertebrates and its outgrowths.- Vital examination.- Preparations in toto.- Examination of sections.- Topographical study.- Morphological details of the epidermal cells.- Histochemical characteristics.- Cutaneous blood vessels.- Cutaneous nerves and tactile receptors.- Skin and hair follicles.- Horny excrescences: nails, horns.- Cutaneous glands.- Tissue blocks containing fragments of bone.- The integument of Arthropods and of other animals having an epidermis covered with a cuticule.- The integument of Molluscs and of other animals with a ciliated epidermis on which glandular cells are scattered.- 32. — Techniques of the histological study of blood and of haematopoietic organs.- Histological examination of the circulating blood.- Blood sampling.- Vital examination.- Permanent preparations.- Hematological stains.- Enumeration of the free elements of the blood.- Histological examination of the haematopoietic organs.- Techniques specially required for the study of haematopoietic organs.- The histological examination of bone marrow.- Histological examination of the spleen.- Histological examination of the thymus.- Histological examination of the lymph nodes and related structures (Tonsils, Peyer patches, in the intestine, etc.).- Histological examination of the haematopoietic organs of invertebrates.- 33. — Techniques of the histological study of the connective tissue.- The histological study of the ground substance.- Histological study of the connective fibres.- Collagen fibres.- Reticulin fibres.- Selective impregnation of reticulin according to Del Rio Hortega.- Silver impregnation according to Gomori.- Silver impregnation according to Oliveira.- Elastic fibres.- Staining with orcein.- Staining with resorcine-fuchsin.- Staining with Gallego’s ferric-fuchsin.- Staining with paraldehyde-fuchsin.- Fibrin.- Fibrinoid.- Oxytalan fibres.- The use of enzymatic preparations for the identification of connective fibres.- Histological study of the connective cells.- 34. — Techniques for the histological study of cartilaginous, bony and dental tissues.- Selective staining of cartilage and of bone on whole mounts.- Histological examination of cartilage.- Techniques specially recommended for the study of chondrocytes.- Techniques particularly recommended for a study of the ground substance.- Techniques specially recommended for the study of fibrillar structures.- Histological examination of bony tissue.- Study of sections of bony tissue prepared by grinding and polishing.- Study of bony tissue on sections taken from fixed material.- Histological examination of dental tissue.- 35. — Techniques for the histological study of muscles and tendons.- Identification of muscle fibres as such.- Demonstration of the structural details of striated muscle fibres.- Demonstration of the structural details of smooth muscle fibres.- Demonstration of the innervation of muscle fibres.- Demonstration of the fibres of tendons.- 36. — Techniques for the histological study of the circulatory apparatus.- Techniques for the histological study of the heart.- Techniques for the histological study of blood vessels.- Methods of following the flow of blood in the tissues and organs.- Techniques which depend on staining the normal content of blood vessels.- Techniques depending on the selective staining of the vascular walls.- Techniques of vascular injection.- 37. — Techniques for the histological study of the digestive apparatus.- Histological examination of the anterior part of the digestive tract.- Histological examination of the stomach.- Histological examination of the intestine.- Gastro-intestinal endocrine cells of the vertebrates:.- Demonstration of all types of endocrine cells by staining with lead haematoxylin.- Methods for selectively demonstrating endocrine cells using silver impregnation.- Methods for the selective demonstration of enterochromamne cells.- Demonstration of theentero-chromamn-like cells or histamine-storing cells.- Demonstration of gastrin cells.- Histological examination of the lymphoidal formations of the digestive tract of vertebrates.- Demonstration of blood vessels and nerves of the gastric and intestinal walls.- Histological examination of the salivary glands.- Histological examination of the exocrine pancreas.- Histological examination of the liver.- Histological examination of the digestive glands of Invertebrates.- The secretory cycle of the glandular formation of the digestive apparatus.- 38. — Techniques for the histological study of the respiratory system.- Histological examination of the respiratory tract of Tetrapods.- Histological examination of the pulmonary tissue.- Histological examination of the swim bladder.- Histological examination of gills.- Histological examination of trachea of Arthropods and related animals.- 39. — Techniques for the histological study of the excretory system.- Histological examination of the kidney of Vertebrates.- Histological examination of the urinary tract of the Vertebrates.- Histological examination of the excretory system of Invertebrates.- 40. — Techniques for the histological study of the nervous system.- Neurohistological Techniques.- Techniques for the detection ofNissl bodies.- Selective coloration of the ganglion cells and their prolongation.- Selective metallic impregnations of the ganglion cells and of their prolongations.- Techniques for neurofibrils.- Nerve-fiber impregnations derived from the silver reduction method.- The osmium-iodide technique for staining nerve-fibres and nerve endings.- Myelin methods.- Production of nerves treated with osmium tetroxide.- Myelin methods derived from Weigert’s method.- Histochemical reactions and diagnostic stagings applicable to the detection of myelin sheaths.- Marchi’s method.- Neuroglia techniques.- Stains for the neuroglia.- Metallic impregnation of neuroglia.- Techniques for the Histological Study of Neurosecretory Cells.- Neurosecretory products which are acidophilic in spite of permanganic oxidation of the sections.- Neurosecretory products which are basophilic after permanganic oxidation.- Recommendations concerning histological techniques applied to the study of the nervous system.- 41. — Techniques for the histological study of sense organs.- Histological examination of the tactile corpuscules.- Histological examination of the pituitary mucosa.- Histological examination of taste buds.- Histological examination of the visual apparatus.- Vital examination of the vertebrate eye.- Histological examination of the eye as a whole.- Methods particularly recommended for the study of different parts of the visual apparatus.- Histological examination of the stato- acoustic apparatus.- 42. — Techniques for the histological study of the endocrine glands.- Technique for the histological study of the neurohaemal organs.- Techniques for the histological study of endocrine glands without accumulation of particular substances.- Techniques for the histological study of endocrine glands which accumulate secretory products.- The distal lobe of the adenohypophysis.- Anatomical study.- Identification of cellular categories.- General histochemical and cytological techniques.- The intermediate lobe of adenohypophysis.- The endocrine pancreas.- Techniques for the quantitative study of the endocrine pancreas.- Techniques for identifying the different types of cell in the endocrine pancreas.- The adrenal gland.- Techniques for the topographical study of the adrenal gland.- Endocrine tissues of Vertebrate gonads.- Thyroid gland.- Epiphysis.- Histological study of hormonal target organs during histophysiological studies on the endocrine glands.- 43. — Techniques for the histological study of the reproductive system.- Histological examination of the male genital system.- Microscopic anatomy of the Vertebrate testes.- Microscopic anatomy of the Invertebrate testes.- The study of spermatogenesis.- The study of the excretory ducts for sperm and of the glands attached to the the male genital system of Vertebrates.- Study of the male genital tracts and of their auxiliary glands in Invertebrates.- Histological study of the female genital system.- Microscopic anatomy of the Vertebrate ovary.- Microscopic anatomy of the female gonads of Invertebrates.- Cytological and histochemical study of ovogenesis.- Histological study of the female genital tracts of Vertebrates.- Histological study of the female genital tracts of Invertebrates.- Appendix IX. — The mass production of histological preparations for teaching purposes.- By way of bibliography.- Index of subjects.