Biological Microtechnique

著者名：Sanderson, Mr Jeremy

Garland Science（2020/08/13発売）

• 言語：ENG
• ISBN：9781872748429
• eISBN：9781000140941
• NDC分類：460.78

Description

A completely new practical guide to both new and classical methods of slide-making which is easy-to-read and easy-to-understand. Biological Microtechnique contains a wealth of practical detail which will provide a firm grounding in preparative methods for light microscopy.

Table of Contents

Abbreviations -- 1. Introduction -- Collecting material for specimen preparation -- Choice of preparative technique -- Looking at preparations -- References -- 2. Fixation -- Function and use of fixatives -- Methods of fixation -- Immersion fixation -- Perfusion fixation -- Vapour fixation -- Phase-partition fixation -- Mechanical methods -- When not to use fixation -- The penetration of fixatives -- Osmolarity and pH -- Fixing agents -- Formaldehyde -- Glutaraldehyde and acrolein -- Alcohols and acetone -- Mercuric chloride -- Potassium dichromate -- Picric acid and acetic acid -- Osmium tetroxide -- Fixative mixtures -- Secondary fixation -- Preservatives -- Washing tissues -- Microwaves in histology -- Microwaving formalin-fixed tissues -- Leiden fixative -- Microwaving paraffin sections -- Microwaving cryostat sections -- Staining reactions -- Safety -- References -- 3. Tissue Processing -- Dehydration -- Transition media -- Processing schedules -- Automatic processing -- Embedding media -- Wax embedding -- Preparation for cutting -- Ribboning -- Ribboning difficulties -- Laying out ribbons -- Mounting sections onto slides -- The water-bath method -- The hot-plate method -- Marking slides -- Locating unstained sections -- Storing wax blocks -- Rehydrating sections -- Gelatin-based embedding media -- Other embedding media -- Polyethylene glycol (PEG) waxes -- Polyester waxes -- Ester wax -- Polyester wax -- Cellulose embedding -- Double embedding -- Orientation of small objects whilst embedding -- Resin embedding media -- Epoxy resins -- Acrylic resins -- Lowicryl and London resins -- Resin removal. -- Methods for hard tissues -- Decalcification -- Determination of end-point -- Sectioning -- Wax impregnation -- Resin impregnation -- Lignified tissues -- Insect tissues -- Hair fibres -- Diatom frustules -- Recording tissue processing -- References -- 4. Microtomy -- Types of microtome -- Hand microtome -- Cambridge rocking microtome -- Rotary microtomes -- Base-sledge and sliding microtomes -- Freezing microtome -- Automated microtomes -- Clamps and chucks -- Types of knives -- Important knife angles and bevels -- Facets -- Care of knives -- Disposable blades -- Glass and diamond knives -- Cryotomy -- Cold knife methods -- Cryostats -- Freezing -- Embedding -- Hazards of cryogenic fluids -- Orientation of tissue -- Cryostat sectioning -- Sectioning temperatures -- Anti-roll plate -- Electrostatic charges -- Handling sections -- Storage of tissue -- Knife sharpening -- Lubricants -- Types of abrasive -- Handles and backs -- Manual sharpening -- Lapping -- Stropping -- Sectioning -- Wax structure -- Compression -- Clearance angle -- Sectioning technique -- Section thickness -- Cryostat sectioning -- Effect of fixation and processing on tissue size -- Sectioning difficulties -- Softening fluids Static -- Summary -- Vibratomes (tissue choppers) -- Macrotomes -- Freehand sectioning References -- 5. Other Preparative Methods -- Cytological methods -- Cytological fixatives -- Cytocentrifuging and sedimentation -- Adherence and loss of cells -- Smears -- Imprints and replicas -- Cell blocks -- Squashes -- Maceration -- Temporary mounts -- Irrigation -- Preparations of whole mounts in cells or 'boxes' -- Dry mounts -- Fluid mounts -- Gum media -- Glycerol jelly mounts -- Glycerol fluid mounts -- References -- 6. Staining and Dyeing -- Nomenclature -- Staining action -- Mordants -- Metachromasia -- Nuclear stains -- Haematoxylin -- Differentiation -- 'Blueing' -- Other nuclear stains -- Carmine -- Safranin -- Synthetic nuclear dyes -- Counterstains -- Eosin -- Other counterstains -- General staining procedures -- Selected staining protocols -- Stains for glycerol jelly mounts -- Stains for resin-embedded material -- Block staining -- Multiple staining of sections -- Cytological stains -- Neutral stains -- Papanicolaou stain -- Polychrome stains -- Vital stains -- Staining for bacteria -- Removal of intrinsic pigments -- Formalin and malarial pigments -- Mercury pigment -- Picric acid -- Osmium tetroxide -- Dye purity -- References -- 7. Finishing the Preparation -- Mountants -- Water-based media -- Dehydration and clearing -- Resinous media -- Mounting technique -- Coverslip thickness -- Adhesives -- Lifting of sections -- Cleaning slides -- Fading of specimens -- Sections stained with fluorochromes -- Finishing the preparation -- Using a ringing table -- Labelling -- Transport -- Artifacts -- Restoring preparations -- Repairing broken slides -- Restaining faded sections -- Restoring tissues dried during processing -- References -- Appendices -- Appendix A: Safety -- Appendix B: Refractive indices -- Index.