Cryo-EM: A Sample preparation and data collection
Gespeichert in:
Format: | Buch |
---|---|
Sprache: | English |
Veröffentlicht: |
Amsterdam [u.a.]
Elsevier, Acad. Press
2010
|
Ausgabe: | 1. ed. |
Schriftenreihe: | Methods in enzymology
481 |
Online-Zugang: | Inhaltsverzeichnis |
Beschreibung: | XLV, 410 S. Ill., graph. Darst. |
ISBN: | 9780123749062 |
Internformat
MARC
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035 | |a (OCoLC)698541319 | ||
035 | |a (DE-599)BVBBV036854541 | ||
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041 | 0 | |a eng | |
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245 | 1 | 0 | |a Cryo-EM |n A |p Sample preparation and data collection |c ed. by Grant J. Jensen |
250 | |a 1. ed. | ||
264 | 1 | |a Amsterdam [u.a.] |b Elsevier, Acad. Press |c 2010 | |
300 | |a XLV, 410 S. |b Ill., graph. Darst. | ||
336 | |b txt |2 rdacontent | ||
337 | |b n |2 rdamedia | ||
338 | |b nc |2 rdacarrier | ||
490 | 1 | |a Methods in enzymology |v 481 | |
490 | 0 | |a Methods in enzymology |v ... | |
700 | 1 | |a Jensen, Grant J. |e Sonstige |4 oth | |
773 | 0 | 8 | |w (DE-604)BV036854506 |g 1 |
830 | 0 | |a Methods in enzymology |v 481 |w (DE-604)BV000000938 |9 481 | |
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943 | 1 | |a oai:aleph.bib-bvb.de:BVB01-020770453 |
Datensatz im Suchindex
_version_ | 1813525205004320768 |
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adam_text |
Contents
Contributors
xi
Preface
xv
Volumes in Series
xvii
3D
Reconstruction from Electron Micrographs:
A Personal Account of its Development
1
David DeRosier
1.
Viruses
2
2.
Helical Structures
7
3. 3D
Reconstruction of Helical Structures
10
4.
Digital Image Processing
12
5. 3D
Reconstruction of Asymmetric Structures
17
References
24
1.
Preparation of 2D Crystals of Membrane Proteins
for High-Resolution Electron Crystallography Data Collection
25
Priyanka D. Abeyrathne,
Mohamed
Chami, Radosav S. Pantelic,
Kenneth
N.
Goldie, and
Henning Stahlberg
1.
Introduction to Electron Crystallography
26
2.
Purification of Membrane Proteins
26
3.
2D Crystallization of Membrane Proteins
28
4.
Requirements for Electron Crystallography Data Collection
32
5.
Conclusions
40
References
41
2.
Helical Crystallization of Soluble and
Membrane Binding Proteins
45
Elizabeth M. Witson-Kubalek, Joshua S. Chappie, and Christopher P. Arthur
1.
Introduction
46
2.
Materials and Methods
49
Acknowledgments
58
References
58
vi
Contents
3.
Plunge Freezing for Electron Cryomicroscopy
63
Megan
J. Dobro,
Linda A. Melanson, Grant J. Jensen,
and Alasdair W. McDowall
1.
Introduction
64
2.
Grids and Supports
65
3.
Cleaning the Grids
66
4.
Preparing the Cryogen
68
5.
Plunging the Grid
71
6.
Common Problems and Their Diagnoses
77
Acknowledgments
78
References
80
4.
A Practical Guide to the Use of Monolayer Purification
and Affinity Grids
83
Deborah F. Kelly,
Danijela Dukovski,
and Thomas
Walz
1.
A Brief History of the Use of
Lipid
Monolayers
in Electron Microscopy
85
2.
Preparation of
Lipid
Monolayer Specimens
86
3.
Monolayer Purification
89
4.
Examples of Monolayer Purification Applications
92
5.
The Affinity Grid
94
6.
Examples of Affinity Grid Applications
98
7.
Characterization of Monolayer Specimens
102
8.
Challenges and Future Directions
104
Acknowledgments
105
References
105
5.
GraFix: Stabilization of Fragile Macromolecular Complexes
for Single Particle Cryo-EM
109
Holger
Stark
1.
Introduction
110
2.
Overview of the GraFix Procedure
112
3.
Methods
120
4.
Conclusions
125
References
125
Contents
vii
6.
Cryonegative Staining of Macromolecular Assemblies
127
Sacha De
Carlo and
Holger
Stark
1.
Introduction
128
2.
Cryonegative Staining—The "Adrian" Method: Frozen-Hydrated
Specimens in the Presence of a Saturated Ammonium Molybdate
Solution at Neutral
pH
133
3.
Cryonegative Staining with the Sandwich Method:
The
"Holger
Stark" Alternative
139
4.
Conclusion
141
Acknowledgments
143
References
143
7.
Liposomes
on a Streptavidin Crystal: A System to
Study Membrane Proteins by Cryo-EM
147
Liguo Wang and Fred J. Sigworth
1.
Introduction
148
2.
Methods
149
3.
Conclusion
162
References
163
8.
Micromanipulator-Assisted Vitreous Cryosectioning
and Sample Preparation by High-Pressure Freezing
165
Mark S. Ladinsky
1.
Introduction
166
2.
Why is Vitreous Cryosectioning so Difficult?
167
3.
High-Pressure Freezing for Vitreous Cryosectioning
168
4.
Extracellular Cryoprotectants for Vitreous Cryosectioning
170
5.
Mounting a Vitrified Sample in the Cryo-Ultramicrotome
171
6.
Hardware: Cryo-Ultramicrotomes
173
7.
The
Micromanipulator 176
8.
Cryodmmond Knives
177
9.
The Cryotrimming Knife
179
10.
Setting up the Cryo-Ultramicrotome for Sectioning
179
11. Trimminga
Biockface
180
12.
Preparing to Cryosection
182
13.
The Active Static Ionizer
183
14.
Cryosectioning
184
15.
Transferring Cryosections to the EM Grid
186
16.
Pressing the Sections onto the Grid
187
17.
The Sectioning Environment and Relative Humidity
189
viii Contents
18.
Cryosectioning
Artifacts
191
19.
Conclusion
192
Acknowledgments
193
References
193
9.
Site-Specific
Biomolecule
Labeling with Gold Clusters
195
Christopher J. Ackerson, Richard D. Powell, and James
F. Hainfeld
1.
Introduction
196
2.
General Considerations
201
3.
Monolayer Protected Cluster Labeling of Biomolecules
203
4.
Nanogold Labeling of Proteins
212
5.
NiOO-NTA-Nanogold Labeling of His-Tagged Proteins
222
6.
Conclusions
225
References
225
10.
How to Operate a Cryo-Electron Microscope
231
Jingchuan Sun and Huilin Li
1.
Introduction
232
2.
Basic Operations of Cryo-EM
233
3.
Several
Practicáis
on Imaging Frozen-Hydrated
Biological Specimens
243
4.
Concluding Remarks
248
References
249
11.
Collecting Electron Crystallographic Data of Two-Dimensional
Protein Crystals
251
Richard
K. Hite,
Andreas D.
Schenk,
Zongli
Li, Yifan Cheng, and Thomas
Walz
1.
Introduction
252
2.
Challenges
253
3.
Images and Diffraction Patterns
255
4.
Specimen Preparation
257
5.
Specimen Flatness
264
6.
Data Collection
266
Acknowledgments
280
References
280
12.
Automated Data Collection for Electron Microscopic Tomography
283
Shawn Q. Zheng, J. W.
Sedat,
and D. A. Agard
1.
Introduction
284
2.
Predictive EMT Data Collection
287
3.
Real-Time
Tomographie
Reconstruction
297
4.
Dual-Axis EMT Data Collection
303
Contents
¡χ
5.
Supporting
Automation
Procedures
309
6.
Summary
312
Acknowledgments
313
References
313
13.
Correlated Light and Electron Cryo-Microscopy
317
Ariane
Briegel,
Songye Chen, Abraham
J.
Koster, Jürgen
M. Plitzko,
Cindi
L.
Schwartz, and Grant
J.
Jensen
1.
Introduction
317
2.
Correlative FLM/ECM with Freezing after FLM Imaging
319
3.
Correlative FLM and ECM, with Freezing before FLM Imaging
322
Acknowledgments
340
References
340
14.
Phase Plates for Transmission Electron Microscopy
343
Radostin Danev and Kuniaki Nagayama
1.
Introduction
344
2.
Types of Phase Plates and Historical Notes
347
3.
Microscope Requirements and Modifications
356
4.
Phase Plate Operation Procedures
359
5.
Summary and Future Prospects
366
Acknowledgments
367
References
367
15.
Radiation Damage in Electron Cryomicroscopy
371
Lindsay A. Baker and John L. Rubinstein
1.
Introduction
372
2.
Measuring Electron Exposure
372
3.
Energy Absorbed by Specimens
374
4.
Radiation Damage and Choice of Accelerating Voltage
374
5.
Primary and Secondary Damage to Proteins During Irradiation
377
6.
Tertiary Damage to Proteins During Irradiation
378
7.
Cryoprotection and Optimal Temperatures
379
8.
Quantification of Beam Damage with Increasing Exposure
381
9.
Optimal Exposures for Thin Crystals
383
10.
Optimal Exposures for Single Particle Samples
384
11.
Optimal Exposures for
Tomographie
Samples
386
12.
Concluding Comments
386
Acknowledgments
387
References
387
Author Index
389
Subject Index
401 |
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illustrated | Illustrated |
indexdate | 2024-10-21T12:06:49Z |
institution | BVB |
isbn | 9780123749062 |
language | English |
oai_aleph_id | oai:aleph.bib-bvb.de:BVB01-020770453 |
oclc_num | 698541319 |
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owner_facet | DE-91G DE-BY-TUM DE-12 DE-355 DE-BY-UBR DE-19 DE-BY-UBM DE-703 |
physical | XLV, 410 S. Ill., graph. Darst. |
publishDate | 2010 |
publishDateSearch | 2010 |
publishDateSort | 2010 |
publisher | Elsevier, Acad. Press |
record_format | marc |
series | Methods in enzymology |
series2 | Methods in enzymology |
spelling | Cryo-EM A Sample preparation and data collection ed. by Grant J. Jensen 1. ed. Amsterdam [u.a.] Elsevier, Acad. Press 2010 XLV, 410 S. Ill., graph. Darst. txt rdacontent n rdamedia nc rdacarrier Methods in enzymology 481 Methods in enzymology ... Jensen, Grant J. Sonstige oth (DE-604)BV036854506 1 Methods in enzymology 481 (DE-604)BV000000938 481 Digitalisierung UB Regensburg application/pdf http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=020770453&sequence=000002&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA Inhaltsverzeichnis |
spellingShingle | Cryo-EM Methods in enzymology |
title | Cryo-EM |
title_auth | Cryo-EM |
title_exact_search | Cryo-EM |
title_full | Cryo-EM A Sample preparation and data collection ed. by Grant J. Jensen |
title_fullStr | Cryo-EM A Sample preparation and data collection ed. by Grant J. Jensen |
title_full_unstemmed | Cryo-EM A Sample preparation and data collection ed. by Grant J. Jensen |
title_short | Cryo-EM |
title_sort | cryo em sample preparation and data collection |
url | http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=020770453&sequence=000002&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA |
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