Biological microtechnique

✍ Scribed by Sanderson, Jeremy B

• Publisher: Bios Scientific Publishing
• Year: 1994
• Tongue: English
• Leaves: 241
• Category: Library

✦ Synopsis

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.;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.

✦ Table of Contents

Cover......Page 1
Half Title......Page 2
Title Page......Page 4
Copyright Page......Page 5
Preface......Page 6
Acknowledgements......Page 8
Table of contents......Page 10
Abbreviations......Page 16
Chapter 1: Introduction......Page 18
Collecting material for specimen preparation......Page 20
Choice of preparative technique......Page 22
Looking at preparations......Page 25
References......Page 27
Function and use of fixatives......Page 28
Immersion fixation......Page 31
Perfusion fixation......Page 32
Phase-partition fixation......Page 33
The penetration of fixatives......Page 34
Osmolarity and pH......Page 35
Formaldehyde......Page 36
Glutaraldehyde and acrolein......Page 37
Potassium dichromate......Page 38
Fixative mixtures......Page 39
Washing tissues......Page 46
Microwaves in histology......Page 47
Microwaving formalin-fixed tissues......Page 49
Microwaving cryostat sections......Page 51
References......Page 52
Chapter 3: Tissue Processing......Page 56
Dehydration......Page 58
Transition media......Page 59
Processing schedules......Page 60
Automatic processing......Page 61
Wax embedding......Page 63
Preparation for cutting......Page 67
Ribboning......Page 68
Mounting sections onto slides......Page 69
The water-bath method......Page 70
The hot-plate method......Page 71
Locating unstained sections......Page 72
Rehydrating sections......Page 73
Gelatin-based embedding media......Page 74
Polyethylene glycol (PEG) waxes......Page 75
Polyester wax......Page 76
Cellulose embedding......Page 77
Double embedding......Page 78
Orientation of small objects whilst embedding......Page 79
Resin embedding media......Page 80
Epoxy resins......Page 82
Acrylic resins......Page 84
Lowicryl and London resins......Page 87
Methods for hard tissues......Page 89
Decalcification......Page 90
Determination of end-point......Page 91
Sectioning......Page 92
Lignified tissues......Page 93
Insect tissues......Page 95
Diatom frustules......Page 96
References......Page 97
Cambridge rocking microtome......Page 102
Rotary microtomes......Page 103
Base-sledge and sliding microtomes......Page 104
Freezing microtome......Page 105
Clamps and chucks......Page 106
Types of knives......Page 107
Important knife angles and bevels......Page 109
Facets......Page 110
Disposable blades......Page 111
Glass and diamond knives......Page 112
Cold knife methods......Page 113
Freezing......Page 114
Hazards of cryogenic fluids......Page 116
Orientation of tissue......Page 117
Cryostat sectioning......Page 118
Electrostatic charges......Page 119
Storage of tissue......Page 121
Knife sharpening......Page 122
Lubricants......Page 123
Handles and backs......Page 124
Manual sharpening......Page 125
Stropping......Page 126
Sectioning......Page 128
Wax structure......Page 129
Compression......Page 130
Sectioning technique......Page 133
Section thickness......Page 136
Effect of fixation and processing on tissue size......Page 137
Sectioning difficulties......Page 140
Static......Page 141
Vibratomes (tissue choppers)......Page 142
Macrotomes......Page 148
Freehand sectioning......Page 149
References......Page 150
Cytological methods......Page 154
Cytological fixatives......Page 155
Cytocentrifuging and sedimentation......Page 156
Adherence and loss of cells......Page 158
Imprints and replicas......Page 159
Cell blocks......Page 161
Maceration......Page 162
Irrigation......Page 163
Preparations of whole mounts in cells or 'boxes'......Page 164
Dry mounts......Page 165
Fluid mounts......Page 168
Glycerol jelly mounts......Page 171
References......Page 174
Nomenclature......Page 176
Staining action......Page 177
Mordants......Page 178
Metachromasia......Page 179
Haematoxylin......Page 180
Other nuclear stains......Page 182
Counterstains......Page 183
General staining procedures......Page 184
Selected staining protocols......Page 187
Stains for glycerol jelly mounts......Page 188
Stains for resin-embedded material......Page 189
Multiple staining of sections......Page 190
Neutral stains......Page 191
Papanicolaou stain......Page 194
Polychrome stains......Page 196
Vital stains......Page 200
Staining for bacteria......Page 201
Formalin and malarial pigments......Page 203
Dye purity......Page 204
References......Page 205
Water-based media......Page 208
Dehydration and clearing......Page 209
Resinous media......Page 210
Mounting technique......Page 211
Coverslip thickness......Page 212
Adhesives......Page 215
Lifting of sections......Page 216
Cleaning slides......Page 217
Fading of specimens......Page 218
Finishing the preparation......Page 219
Using a ringing table......Page 220
Transport......Page 222
Artifacts......Page 224
Repairing broken slides......Page 226
Restoring tissues dried during processing......Page 227
References......Page 228
Appendix A: Safety......Page 230
Appendix B: Refractive indices......Page 234
Index......Page 236

✦ Subjects

Microtechnique;SCIENCE / Life Sciences / General;Electronic books