Introduction to electron microscopy for biologists:
Saved in:
| Format: | Book |
|---|---|
| Language: | English |
| Published: |
Amsterdam [u.a.]
Elsevier, Academic Press
2008
|
| Edition: | 1. ed. |
| Series: | Methods in cell biology
88 |
| Subjects: | |
| Online Access: | Inhaltsverzeichnis |
| Physical Description: | XXII, 545 S. Ill., graph. Darst. |
| ISBN: | 9780123743206 |
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MARC
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| 245 | 1 | 0 | |a Introduction to electron microscopy for biologists |c ed. by Terence D. Allen |
| 250 | |a 1. ed. | ||
| 264 | 1 | |a Amsterdam [u.a.] |b Elsevier, Academic Press |c 2008 | |
| 300 | |a XXII, 545 S. |b Ill., graph. Darst. | ||
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| 490 | 1 | |a Methods in cell biology |v 88 | |
| 650 | 7 | |a Cytologie - Technique |2 ram | |
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| 650 | 4 | |a Cytology |x Technique | |
| 650 | 4 | |a Electron microscopy | |
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Record in the Search Index
| _version_ | 1804137962622943232 |
|---|---|
| adam_text | CONTENTS
Contributors
Preface
PART I Exploring the Organisation of the Cell by
Electron Microscopy
SECTION
1
Basic Transmission and Scanning Electron Microscopy
1.
High Pressure Freezing and Freeze Substitution of Schizosaccharomyces pombe
and Saccharomyces cerevisiae for
ТЕМ
Stephen Murray
I. Introduction
4
II. Materials and Instrumentation
4
III. Procedures
7
IV. Comments and Problems
10
References
17
2.
Electron Probe X-ray Microanalysis for the Study of Cell Physiology
E.
Fernandez-Segura
and Alice Warley
I. Introduction
20
II. Rationale
21
III. Methods
22
IV. Equipment
37
V. Discussion
38
References
40
3.
Preparation of Cells and Tissues for Immuno EM
Paul Webster, Heinz
Schwarz,
and
Garelli
Griffiths
I. Preparing Biological Specimens for Examination by Electron Microscopy
46
II. Vitrification and Chemical Fixation for Immunolocalization
48
Contents
III.
Vitrification
Followed by Freeze Substitution
48
IV. Chemical Cross-linking (or Fixation)
49
V. Embedding in Resin for Sectioning
50
VI. Cryosectioning for Immunocytochemistry: The Tokuyasu Method
50
VII.
The Starting Material
51
VIII.
Protocols
51
IX. Alternative Approaches to Freeze Substitution
53
X. Correlative Microscopy
54
References
55
4.
Quantification of Structures and Gold Labeling in Transmission Electron Microscopy
John Lucocq
I. Introduction
59
II. Sampling and Stereology
60
III. Quantities Displayed on Sections
—
Gold Labeling and Profile Data
62
IV. Quantities in Three Dimension
69
V. Spatial Analysis
79
References
80
5.
Combined Video Fluorescence and
3D
Electron Microscopy
Alexander A, Mironov,
Koman
S.
Poìishchuk,
and
Galina V. Beznoussenko
I. Introduction
84
II. Rationale
84
III. Method Steps
85
IV. Immunolabeling for EM with NANOGOLD
87
V. Immunolabeling for EM with
HRP 88
References
95
6.
From Live-Cell Imaging to Scanning Electron Microscopy
(SEM):
The Use
of Green Fluorescent Protein (GFP) as a Common Label
Sheona P.
Drummond
and Terence D. Allen
I. Introduction
98
II. Rationale
99
III. Methods
99
IV. Summary
103
V. Concluding Remarks
104
References
107
Contents
7. Immunolabeling
for Scanning
Electron
Microscopy
(SEM)
and Field Emission
SEM
Martin
W.
Goldberg
I. Introduction HO
II. Methods
120
III. Procedure
122
Conclusions
129
References
129
Immunogold Labeling of Thawed Cryosections
Peter J. Peters and Jason Pierson
I. Introduction
132
II. Preparation of Carbon-and Formvar-Coated Copper Girds
132
III. Aldehyde Fixation of Cells for Cryoimmuno Labeling
134
IV. Embedding Samples for Cryoimmuno Labeling
136
V. Cryosectioning for Cryoimmuno Labeling
138
VI. Immunogold Labeling
140
VII.
Reagents and Solutions
143
VIII.
Future Outlooks
146
References
148
9.
Close-to-Native
Ultrastructural
Preservation by High Pressure Freezing
Dimitri Vanhecke, Werner
Graber,
and Daniel
Stader
I. Introduction
152
II. Rational
155
III. Material
156
IV. Methods
158
V. High Pressure Frozen Samples
159
VI. Discussion
160
References
162
10.
High-Pressure Freezing and Low-Temperature Fixation of Cell Monolayers
Grown on Sapphire
Coverslips
Siegfried Reipert and Gerhard
Wiche
I. Introduction
166
II. Materials and Instrumentation
167
III. Procedures
167
IV. Comments and
Pitfaîîs
174
References
180
Contents
11. Freeze-Fracture
Cytochemistry in Cell Biology
Nicholas
J.
Severs and
Horst
Robenek
I. Introduction
182
II. Basic Rationale: Solving the Problems of Combining Freeze
Fracture with Cytochemistry
183
III. Methods
185
IV. Discussion: Impact on Topical Questions in Cell Biology
191
V. Concluding Comment
201
References
202
PART II Electron Microscopy of Specific Cellular Structure
SECTION
1
The Cell Membrane
12.
Three-Dimensional Molecular Architecture of the Plasma-Membrane-Associated
Cytoskeleton as Reconstructed by Freeze-Etch Electron Tomography
Nobuhiro
Moroni ,
Chieko Nakada, Yasuhiro Umemura,
Jiro
Usukura, and Akihiro Kusumi
I. Introduction
208
II. Protocol for Visualization of the Three-Dimensional Structure of the
MSK
of the Cytoplasmic Surface of the Plasma Membrane
212
III.
3D
Structure of the Cytoskeleton-Plasma Membrane Interface
219
IV. Electron Tomography Clarified that Some of the Actin Filaments are
Laterally Bound to the Cytoplasmic Surface of the Plasma Membrane
231
References
232
13.
Visualization of
Dynamins
Jason
Λ.
Hears and Jenny
E.
Hinshaw
I. Introduction
237
II. Methods and Materials
240
III. Discussion
251
IV. Summary
253
References
254
Contents
SECTION
2
The Cytoskeleton
14.
Correlated Light and Electron Microscopy of the Cytoskeleton
Sonja Auinger
ana]. Victor Small
I. Introduction
258
II. Materials and Methods
259
III. Methods
260
IV. Results and Discussion
266
V. Summary
271
References
271
15.
Electron Microscopy of Intermediate Filaments: Teaming up with
Atomic Force and Confocal Laser Scanning Microscopy
Laurent Kreplak, Karsten Richter, Ueli Aebi,
and
Harald
Herrmann
I. Introduction
274
II. Rationale
277
III. Visualization of Intermediate Filaments in vitro and in Cultured Cells
278
IV. Materials
293
V. Conclusions and Outlook
294
References
295
16.
Studying Microtubules by Electron Microscopy
Carolyn
Moores
I. Introduction
300
II. Molecular Electron Microscopy
302
III. Cellular Electron Microscopy
311
IV. Outlook
314
References
315
SECTION
3
Extracellular Matrix and Cell Junctions
17.
Electron Microscopy of Collagen Fibril Structure In Vitro and In Vivo Including
Three-Dimensional Reconstruction
Tobias
Starborg,
Yinhui
Lu,
Karl E. Kadler, and David P. Holmes
I. Introduction
320
II. Electron Microscopy of Isolated Collagen Fibrils
327
III. Fibroblast/Fibril Interface in Developing Tendon
334
IV. Discussion
340
V. Conclusions
341
References
341
Contents
18.
Visualization of Desmosomes in the Electron Microscope
Anthea Scothern and David Garrod
I. Introduction
348
II. Rationale
351
III. Methods
351
IV. Materials
360
V. Discussion
364
VI. Summary
364
References
365
SECTION
4
The Nucleus
19.
A Protocol for Isolation and Visualization of Yeast Nuclei by Scanning
Electron Microscopy
Stephen Munay and Elena Kiseleva
I. Introduction
368
II. Materials and Instrumentation
370
HI. Procedures
372
IV. Comments and Problems
385
References
386
20.
Scanning Electron Microscopy of Nuclear Structure
Terence D. Allen, Sandra A
.
Rutherford, Stephen Murray, Sheona P.
Drummond,
Martin W. Goldberg, and Elena Kiseleva
I. Introduction
390
II. Rationale
393
HI. Methods
393
IV. Colloidal Gold in the
SEM 400
V. Immunolabeling Protocol
403
VI. CPD for High Resolution
SEM 404
VII.
Discussion
407
References
408
21.
Electron Microscopy of
Lamin
and the Nuclear Lamina in Caenorhabditis
elegans
Merau Cohen, Rachel Santarella, Naama
Wiesel,
lain Mattaj, and Yosef Gruenbaum
I. General Introduction
412
II. In Vitro Assembly of Ce-Lamin Filaments
414
HI. Preparation of Embryos and Adults for Transmission Electron
Microscopy Using Microwave Fixation
416
Contents
IV. Preparation of C.
elegans Embryos
and Adults for Conventional
Transmission Electron
Microscopy by High Pressure Freezing
Combined with Freeze Substitution
419
V. Preembedding Immunogold EM Staining of Lamina Proteins
in
C. elegans
Embryos
423
VI. Postembedding Immunogold EM Staining of Lamina Proteins
in
C. elegans
Embryos
425
VII.
Summary
427
References
427
22.
Visualization of Nuclear Organization by Ultrastructural Cytochemistry
Marco Biggiogem and
Stanislav Fakan
I. Introduction
432
II. Cytochemical Contrasting Approaches
432
III. High Resolution Autoradiography
441
IV. Immunocytochemistry
441
V. Molecular in situ Hybridization
443
VI. Identification of Nucleic Acids by Means of Enzymatic Reactions
444
VII.
Targeting of Intranuclear Substrates using Enzyme-Colloidal
Gold Complexes
446
VIII.
Concluding Remarks
446
References
447
23.
Scanning Electron Microscopy of Chromosomes
Gerhard Wanner and Elizabeth Schroeder-Reiter
I. Introduction
452
II. Materials and Methods
454
III. Chromosome Preparation
456
IV. Chromosome Structure in
SEM
458
V. Chromosome Analysis in
SEM
460
VI. Outlook
470
References
473
PART III Cells and Infectious Agents
24.
Infection at the Cellular Level
Christian Goosmann,
Ulrike
Abu Abed, and
Volker Brinkmann
I. Introduction
477
II. Methods and Materials
478
III. Overview and Conclusion
494
References
495
Contents
25. Electron
Microscopy of Viruses and Virus-Cell Interactions
Peter Wild
I. Introduction
498
II. Methods
500
III. Material
515
IV. Discussion
516
V. Summary
520
References
521
Index
525
Volumes in Series
537
|
| adam_txt |
CONTENTS
Contributors
Preface
PART I Exploring the Organisation of the Cell by
Electron Microscopy
SECTION
1
Basic Transmission and Scanning Electron Microscopy
1.
High Pressure Freezing and Freeze Substitution of Schizosaccharomyces pombe
and Saccharomyces cerevisiae for
ТЕМ
Stephen Murray
I. Introduction
4
II. Materials and Instrumentation
4
III. Procedures
7
IV. Comments and Problems
10
References
17
2.
Electron Probe X-ray Microanalysis for the Study of Cell Physiology
E.
Fernandez-Segura
and Alice Warley
I. Introduction
20
II. Rationale
21
III. Methods
22
IV. Equipment
37
V. Discussion
38
References
40
3.
Preparation of Cells and Tissues for Immuno EM
Paul Webster, Heinz
Schwarz,
and
Garelli
Griffiths
I. Preparing Biological Specimens for Examination by Electron Microscopy
46
II. Vitrification and Chemical Fixation for Immunolocalization
48
Contents
III.
Vitrification
Followed by Freeze Substitution
48
IV. Chemical Cross-linking (or Fixation)
49
V. Embedding in Resin for Sectioning
50
VI. Cryosectioning for Immunocytochemistry: The Tokuyasu Method
50
VII.
The Starting Material
51
VIII.
Protocols
51
IX. Alternative Approaches to Freeze Substitution
53
X. Correlative Microscopy
54
References
55
4.
Quantification of Structures and Gold Labeling in Transmission Electron Microscopy
John Lucocq
I. Introduction
59
II. Sampling and Stereology
60
III. Quantities Displayed on Sections
—
Gold Labeling and Profile Data
62
IV. Quantities in Three Dimension
69
V. Spatial Analysis
79
References
80
5.
Combined Video Fluorescence and
3D
Electron Microscopy
Alexander A, Mironov,
Koman
S.
Poìishchuk,
and
Galina V. Beznoussenko
I. Introduction
84
II. Rationale
84
III. Method Steps
85
IV. Immunolabeling for EM with NANOGOLD
87
V. Immunolabeling for EM with
HRP 88
References
95
6.
From Live-Cell Imaging to Scanning Electron Microscopy
(SEM):
The Use
of Green Fluorescent Protein (GFP) as a Common Label
Sheona P.
Drummond
and Terence D. Allen
I. Introduction
98
II. Rationale
99
III. Methods
99
IV. Summary
103
V. Concluding Remarks
104
References
107
Contents
7. Immunolabeling
for Scanning
Electron
Microscopy
(SEM)
and Field Emission
SEM
Martin
W.
Goldberg
I. Introduction HO
II. Methods
120
III. Procedure
122
Conclusions
129
References
129
Immunogold Labeling of Thawed Cryosections
Peter J. Peters and Jason Pierson
I. Introduction
132
II. Preparation of Carbon-and Formvar-Coated Copper Girds
132
III. Aldehyde Fixation of Cells for Cryoimmuno Labeling
134
IV. Embedding Samples for Cryoimmuno Labeling
136
V. Cryosectioning for Cryoimmuno Labeling
138
VI. Immunogold Labeling
140
VII.
Reagents and Solutions
143
VIII.
Future Outlooks
146
References
148
9.
Close-to-Native
Ultrastructural
Preservation by High Pressure Freezing
Dimitri Vanhecke, Werner
Graber,
and Daniel
Stader
I. Introduction
152
II. Rational
155
III. Material
156
IV. Methods
158
V. High Pressure Frozen Samples
159
VI. Discussion
160
References
162
10.
High-Pressure Freezing and Low-Temperature Fixation of Cell Monolayers
Grown on Sapphire
Coverslips
Siegfried Reipert and Gerhard
Wiche
I. Introduction
166
II. Materials and Instrumentation
167
III. Procedures
167
IV. Comments and
Pitfaîîs
174
References
180
Contents
11. Freeze-Fracture
Cytochemistry in Cell Biology
Nicholas
J.
Severs and
Horst
Robenek
I. Introduction
182
II. Basic Rationale: Solving the Problems of Combining Freeze
Fracture with Cytochemistry
183
III. Methods
185
IV. Discussion: Impact on Topical Questions in Cell Biology
191
V. Concluding Comment
201
References
202
PART II Electron Microscopy of Specific Cellular Structure
SECTION
1
The Cell Membrane
12.
Three-Dimensional Molecular Architecture of the Plasma-Membrane-Associated
Cytoskeleton as Reconstructed by Freeze-Etch Electron Tomography
Nobuhiro
Moroni',
Chieko Nakada, Yasuhiro Umemura,
Jiro
Usukura, and Akihiro Kusumi
I. Introduction
208
II. Protocol for Visualization of the Three-Dimensional Structure of the
MSK
of the Cytoplasmic Surface of the Plasma Membrane
212
III.
3D
Structure of the Cytoskeleton-Plasma Membrane Interface
219
IV. Electron Tomography Clarified that Some of the Actin Filaments are
Laterally Bound to the Cytoplasmic Surface of the Plasma Membrane
231
References
232
13.
Visualization of
Dynamins
Jason
Λ.
Hears and Jenny
E.
Hinshaw
I. Introduction
237
II. Methods and Materials
240
III. Discussion
251
IV. Summary
253
References
254
Contents
SECTION
2
The Cytoskeleton
14.
Correlated Light and Electron Microscopy of the Cytoskeleton
Sonja Auinger
ana]. Victor Small
I. Introduction
258
II. Materials and Methods
259
III. Methods
260
IV. Results and Discussion
266
V. Summary
271
References
271
15.
Electron Microscopy of Intermediate Filaments: Teaming up with
Atomic Force and Confocal Laser Scanning Microscopy
Laurent Kreplak, Karsten Richter, Ueli Aebi,
and
Harald
Herrmann
I. Introduction
274
II. Rationale
277
III. Visualization of Intermediate Filaments in vitro and in Cultured Cells
278
IV. Materials
293
V. Conclusions and Outlook
294
References
295
16.
Studying Microtubules by Electron Microscopy
Carolyn
Moores
I. Introduction
300
II. Molecular Electron Microscopy
302
III. Cellular Electron Microscopy
311
IV. Outlook
314
References
315
SECTION
3
Extracellular Matrix and Cell Junctions
17.
Electron Microscopy of Collagen Fibril Structure In Vitro and In Vivo Including
Three-Dimensional Reconstruction
Tobias
Starborg,
Yinhui
Lu,
Karl E. Kadler, and David P. Holmes
I. Introduction
320
II. Electron Microscopy of Isolated Collagen Fibrils
327
III. Fibroblast/Fibril Interface in Developing Tendon
334
IV. Discussion
340
V. Conclusions
341
References
341
Contents
18.
Visualization of Desmosomes in the Electron Microscope
Anthea Scothern and David Garrod
I. Introduction
348
II. Rationale
351
III. Methods
351
IV. Materials
360
V. Discussion
364
VI. Summary
364
References
365
SECTION
4
The Nucleus
19.
A Protocol for Isolation and Visualization of Yeast Nuclei by Scanning
Electron Microscopy
Stephen Munay and Elena Kiseleva
I. Introduction
368
II. Materials and Instrumentation
370
HI. Procedures
372
IV. Comments and Problems
385
References
386
20.
Scanning Electron Microscopy of Nuclear Structure
Terence D. Allen, Sandra A
.
Rutherford, Stephen Murray, Sheona P.
Drummond,
Martin W. Goldberg, and Elena Kiseleva
I. Introduction
390
II. Rationale
393
HI. Methods
393
IV. Colloidal Gold in the
SEM 400
V. Immunolabeling Protocol
403
VI. CPD for High Resolution
SEM 404
VII.
Discussion
407
References
408
21.
Electron Microscopy of
Lamin
and the Nuclear Lamina in Caenorhabditis
elegans
Merau Cohen, Rachel Santarella, Naama
Wiesel,
lain Mattaj, and Yosef Gruenbaum
I. General Introduction
412
II. In Vitro Assembly of Ce-Lamin Filaments
414
HI. Preparation of Embryos and Adults for Transmission Electron
Microscopy Using Microwave Fixation
416
Contents
IV. Preparation of C.
elegans Embryos
and Adults for Conventional
Transmission Electron
Microscopy by High Pressure Freezing
Combined with Freeze Substitution
419
V. Preembedding Immunogold EM Staining of Lamina Proteins
in
C. elegans
Embryos
423
VI. Postembedding Immunogold EM Staining of Lamina Proteins
in
C. elegans
Embryos
425
VII.
Summary
427
References
427
22.
Visualization of Nuclear Organization by Ultrastructural Cytochemistry
Marco Biggiogem and
Stanislav Fakan
I. Introduction
432
II. Cytochemical Contrasting Approaches
432
III. High Resolution Autoradiography
441
IV. Immunocytochemistry
441
V. Molecular in situ Hybridization
443
VI. Identification of Nucleic Acids by Means of Enzymatic Reactions
444
VII.
Targeting of Intranuclear Substrates using Enzyme-Colloidal
Gold Complexes
446
VIII.
Concluding Remarks
446
References
447
23.
Scanning Electron Microscopy of Chromosomes
Gerhard Wanner and Elizabeth Schroeder-Reiter
I. Introduction
452
II. Materials and Methods
454
III. Chromosome Preparation
456
IV. Chromosome Structure in
SEM
458
V. Chromosome Analysis in
SEM
460
VI. Outlook
470
References
473
PART III Cells and Infectious Agents
24.
Infection at the Cellular Level
Christian Goosmann,
Ulrike
Abu Abed, and
Volker Brinkmann
I. Introduction
477
II. Methods and Materials
478
III. Overview and Conclusion
494
References
495
Contents
25. Electron
Microscopy of Viruses and Virus-Cell Interactions
Peter Wild
I. Introduction
498
II. Methods
500
III. Material
515
IV. Discussion
516
V. Summary
520
References
521
Index
525
Volumes in Series
537 |
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| id | DE-604.BV035031777 |
| illustrated | Illustrated |
| index_date | 2024-07-02T21:50:06Z |
| indexdate | 2024-07-09T21:20:38Z |
| institution | BVB |
| isbn | 9780123743206 |
| language | English |
| oai_aleph_id | oai:aleph.bib-bvb.de:BVB01-016700751 |
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| physical | XXII, 545 S. Ill., graph. Darst. |
| publishDate | 2008 |
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| publisher | Elsevier, Academic Press |
| record_format | marc |
| series | Methods in cell biology |
| series2 | Methods in cell biology |
| spelling | Introduction to electron microscopy for biologists ed. by Terence D. Allen 1. ed. Amsterdam [u.a.] Elsevier, Academic Press 2008 XXII, 545 S. Ill., graph. Darst. txt rdacontent n rdamedia nc rdacarrier Methods in cell biology 88 Cytologie - Technique ram Microscopie électronique ram Cytology Technique Electron microscopy Neophyten Botanik (DE-588)4271854-5 gnd rswk-swf Elektronenmikroskopie (DE-588)4014327-2 gnd rswk-swf Zelle (DE-588)4067537-3 gnd rswk-swf Britische Inseln (DE-588)4090131-2 gnd rswk-swf (DE-588)4143413-4 Aufsatzsammlung gnd-content Zelle (DE-588)4067537-3 s Elektronenmikroskopie (DE-588)4014327-2 s DE-604 Britische Inseln (DE-588)4090131-2 g Neophyten Botanik (DE-588)4271854-5 s b DE-604 Allen, Terence 1944- Sonstige (DE-588)136680674 oth Methods in cell biology 88 (DE-604)BV002534878 88 Digitalisierung UB Regensburg application/pdf http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=016700751&sequence=000002&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA Inhaltsverzeichnis |
| spellingShingle | Introduction to electron microscopy for biologists Methods in cell biology Cytologie - Technique ram Microscopie électronique ram Cytology Technique Electron microscopy Neophyten Botanik (DE-588)4271854-5 gnd Elektronenmikroskopie (DE-588)4014327-2 gnd Zelle (DE-588)4067537-3 gnd |
| subject_GND | (DE-588)4271854-5 (DE-588)4014327-2 (DE-588)4067537-3 (DE-588)4090131-2 (DE-588)4143413-4 |
| title | Introduction to electron microscopy for biologists |
| title_auth | Introduction to electron microscopy for biologists |
| title_exact_search | Introduction to electron microscopy for biologists |
| title_exact_search_txtP | Introduction to electron microscopy for biologists |
| title_full | Introduction to electron microscopy for biologists ed. by Terence D. Allen |
| title_fullStr | Introduction to electron microscopy for biologists ed. by Terence D. Allen |
| title_full_unstemmed | Introduction to electron microscopy for biologists ed. by Terence D. Allen |
| title_short | Introduction to electron microscopy for biologists |
| title_sort | introduction to electron microscopy for biologists |
| topic | Cytologie - Technique ram Microscopie électronique ram Cytology Technique Electron microscopy Neophyten Botanik (DE-588)4271854-5 gnd Elektronenmikroskopie (DE-588)4014327-2 gnd Zelle (DE-588)4067537-3 gnd |
| topic_facet | Cytologie - Technique Microscopie électronique Cytology Technique Electron microscopy Neophyten Botanik Elektronenmikroskopie Zelle Britische Inseln Aufsatzsammlung |
| url | http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=016700751&sequence=000002&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA |
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| work_keys_str_mv | AT allenterence introductiontoelectronmicroscopyforbiologists |