Biological Microtechnique

ISBN-13: 9781872748429 / Angielski / Miękka / 1994 / 240 str.

J. B. Sanderson; Jeremy B. Sanderson; Dan Sanderson

Opis

Although many significant advances have been made in biological specimen preparation during the past 20 years no new practical guide to the techniques has been published in this time. As a result of the recent resurgence of interest in light microscopy, particularly confocal techniques, this up-to-date book should benefit both the professional and amateur alike.

Recenzje

'... little of consequence is omitted, and virtually all of real use is present. Significant advances of the last 20 years are covered in as much detail as most will ever need ... an admirable volume.' - Journal of Microscopy

Spis treści

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.

O autorze

Jeremy Sanderson