A Manual of Applied Techniques for Biological Electron Microscopy

Front Cover
Springer Science & Business Media, Aug 31, 1993 - Science - 257 pages
0 Reviews
This easy-to-follow manual describes tested procedures used to prepare biological samples for scanning and transmission electron microscopy, as well as methods for cytochemistry, immunocytochemistry, and scientific photography. The work is structured to clearly define testing objectives, necessary materials, procedural steps, and expected results; a list of references and trouble shooting techniques round out the text.
 

What people are saying - Write a review

We haven't found any reviews in the usual places.

Contents

Specimen Preparation
1
Physical Fixation
5
EMBEDDING MEDIA
6
MISCELLANEOUS TIPS FOR FORMULATING SOLUTIONS IN THE ELECTRON MICROSCOPY LABORATORY
9
A Routine Fixation and Embedding Schedule for Transmission Electron Microscopy Samples Tissues or Cells
12
A Routine Schedule for the Preparation of Cell and Tissue Samples for Scanning Electron Microscopy
18
Preparation of Primary Aldehyde Fixatives
20
Preparation of Osmium Tetroxide
22
Semithin Section Staining with Toluidine Blue O
120
Polychrome Stain for Semithin Sections
122
Staining Ultrathin Sections
123
Support Films
129
FormvarCoated Aluminum Bridges for Slot Grids
133
Coating Grids with Butvar B98
139
Coating Grids with Collodion Films
141
Making Carbon Support Films
143

BUFFERING SYSTEMS
28
Cacodylate Buffers
29
Phosphate Buffers
30
TrisHCI Buffer
32
Sodium Acetate Buffer
33
Dulbeccos PhosphateBuffered Saline
34
RESIN FORMULATIONS
36
Spurr Resin
38
PolyBed 812 Resin
42
SPIPON 812 Kit
44
Epon 812
46
Araldite 6005 Resin
47
Mollenhauers EponAraldite Resin
49
LR London Resin Co White Resin
50
Lowicryl and LR Gold Resins
52
Polyethylene Glycol Method for TEM Sections
53
JB4 Glycol Methacrylate Techniques for HighResolution Light Microscopy
55
Agar Embedment of Cell Suspensions or Subcellular Particulates for TEM
57
Preparing Adherent Tissue Culture Monolayers in Situ for TEM
60
Flat Embedding of Cell Cultures Grown on Permanox Tissue Culture Dishes for TEM
63
Preparation of Buffy Coats for TEM
64
Sperm Fixation
65
Central Nervous System Fixation Brain Spinal Cord
67
Using Vacuum to Help Wet Fungal Plant or Insect Materials during Primary Fixation
69
Simultaneous Aldehyde Osmium Fixation for Protozoan Samples or Samples with Large Amounts of Lipid
70
Killing Cells Prior to Chemical Fixation
72
Flat Embedding on Microscope Slides
73
Procedure for Deparaffining Tissues
75
Sectioning Methods
79
Making a Section Manipulation Tool
82
Making a Locking Ring for Forceps
83
Making Glass Knife Boats
85
Glass Knife Storage
87
Diamond Knives
88
Block Trimming
89
Sectioning Procedures
90
Semithin Sections
95
Grid Selection
96
Grid Cleaning
97
Common Sectioning Problems
98
Specialized SEM Procedures
101
PolyLlysine Technique for Attaching Particulates to Coverslips for SEM
102
Organosilane CoverslipSlide Treatment
104
Critical Point Drying for SEM
106
Drying Samples with Hexamethyldisilazane
109
Drying SEM Samples with Peldri II
110
Sputter Coating
111
Vascular Casting with Mercox CL2B
114
Section Staining Procedures
119
Negative Staining
147
Negative Staining with Phosphotungstic Acid
148
Negative Staining with Uranyl Acetate
151
Negative Staining with Ammonium Molybdate
154
Wetting Agents Used for Negative Staining
155
Preparation of Viral Samples for Transmission Electron Microscopy
159
Lyphogel Technique for Concentrating Virus
161
Immune Electron Microscopy for Concentrating Virus
162
Concentrating Virus with the Beckman Airfuge
163
Vacuum Evaporation
167
Shadow Casting
170
DNA Plasmid Preparation for TEM
174
Films and Photographic Methods
177
Kodak Electron Microscope TEM Film 4489
180
Films in 35mm Format
181
Kodak Technical Pan 2415 for Photomicrography
183
Ilford PanF and FP4
185
Kodak TMax 100 Film
186
Kodalith Film
187
Kodak LPD4Precision Line Film
188
Kodak Rapid Process Copy Film 2064
189
Kodak Recordak 2468
190
Polaroid Polagraph HC 35mm Film
191
Polaroid Polaplan 35mm Film
192
Kodak Technical Pan 2415 for Making Copy Negatives
194
Polaroid Copy Negatives Using Type 55 PN Film
195
Making Photographic Prints
196
Poster Preparation
200
Cytochemical Techniques
205
POLYSACCHARIDE STAINS
206
Ruthenium Red Staining
207
Silver Methenamine Staining for Polysaccharides
208
CALCIUM STAINING
213
Postfixation Calcium Staining with Pyroantimonate
216
COLLOIDAL GOLD TECHNIQUES
218
Preparation of 13nm Colloidal Gold
220
Conjugation of Gold to Proteins
223
Indirect Immunolabeling Procedure for Sections of Materials Embedded in LR White or Lowicryl K4M Resin Acrylic Resins
226
Procedure for Immunolabeling Intact Cells Preembedment Labeling of Cell Surfaces
228
Cryoultramicrotomy and Subsequent Immunolabeling an Example for Muscle Immunolabeling
229
Autoradiography
235
Autoradiography with Semithin EpoxideEmbedded Sections of Cells or Tissues
236
Computing Micrometer Bar Sizes
243
Calibrating the TEM and the SEM
245
Materials and Methods Writeup Suggestions for Standard TEM and SEM Preparations
247
Safety
249
Electron Microscopy Supplies
251
Index
253
Copyright

Common terms and phrases

References to this book

All Book Search results »

Bibliographic information