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Tables of Contents for Methods in Cell Biology
Chapter/Section Title
Page #
Page Count
Contributors
ix
Preface
xi
Introduction to Confocal Microscopy
Shirley J. Wright
David J. Wright
Introduction
2
5
Principle of Confocal Microscopy
7
7
Major Components of a Confocal Microscope
14
9
Designs of Confocal Microscopes
23
26
Choosing and Setting Up a Confocal Microscope System
49
3
Specimen Preparation
52
1
Applications of Confocal Microscopy
52
16
Alternatives to Confocal Microscopy
68
5
Conclusions and Future Directions
73
1
Resources for Confocal Microscopy
74
14
References
81
7
Direct-View High-Speed Confocal Scanner: The CSU-10
Shinya Inoue
Ted Inoue
Introduction
88
2
Overview of Confocal Microscopes
90
1
Design of the CSU-10
91
2
Sample Biological Applications
93
8
Low Bleaching of Fluorescence Observed with the CSU-10
101
3
Very-High-Speed Full-Frame Confocal Imaging with the CSU-10
104
1
Haze Removal, Image Sharpening, and Dynamic Stereo Image Generation by Digital Signal Processing
105
12
Mechanical and Optical Performance of the CSU-10
117
1
Potentials of the CSU-10
118
4
Addendum A (April, 2002)
122
2
Addendum B (personal communication from Dr. Kenneth R. Spring, June, 2002)
124
5
References
125
4
Introduction to Multiphoton Excitation Imaging for the Biological Sciences
Victoria E. Centonze
Introduction
129
2
Principles of Multiphoton Imaging
131
2
Advantages of Multiphoton Imaging
133
4
Imaging Parameters
137
4
Applications of Multiphoton Imaging
141
3
New Development in Nonlinear Optics
144
1
Conclusion
145
4
References
145
4
Confocal Microscopy: Important Considerations for Accurate Imaging
Lars Majlof
Per-Ola Forsgren
Introduction
149
1
Factors Affecting Confocal Imaging
150
13
Conclusions
163
3
References
164
2
Multicolor Laser Scanning Confocal Immunofluorescence Microscopy: Practical Application and Limitations
T. Clark Brelje
Martin W. Wessendorf
Robert L. Sorenson
Introduction
166
1
Immunofluorescence Histochemistry
167
18
Instrumentation
185
6
Approaches to Multicolor Laser Scanning Confocal Microscopy
191
16
Practical Aspects of Multicolor Laser Scanning Confocal Microscopy
207
26
Multicolor Laser Scanning Confocal Microscopy Considerations
233
7
Conclusions
240
7
References
241
6
Practical Aspects of Objective Lens Selection for Confocal and Multiphoton Digital Imaging Techniques
Gerald S. Benham
Introduction
247
2
Confocal Microscopy Resolution
249
5
Infinity versus Finite Lens Systems
254
2
Objective Lens Nomenclature and Features
256
5
Objective Lens Aberrations
261
2
Dry (Nonimmersion) and Immersion Objective Lenses
263
3
Epi (Reflected Light) Objectives
266
2
Live Cell and Fixed Tissue Confocal Applications and Objective Lens Selection
268
10
Possible Solutions for Potential Problematic Applications
278
10
Optimum Objective Lenses for Multiphoton Applications
288
1
Transmission Data for Confocal and Multiphoton Objectives
289
1
Conclusions
290
12
References
293
9
Resolution of Subcellular Detail in Thick Tissue Sections: Immunohistochemical Preparation and Fluorescence Confocal Microscopy
Irene L. Hale
Brian Matsumoto
Introduction: Development of Fluorescence Microscopy Techniques for Resolution of Subcellular Detail
302
6
Immunohistochemical Preparation of Thick Tissue Sections
308
13
Confocal Imaging Parameters for Maximum Resolution
321
5
Applications
326
3
Appendix: Procedures for Immunohistochemical Preparation
329
9
References
332
6
Confocal Fluorescence Microscopy for Tetrahymena thermophila
E. S. Cole
K. R. Stuart
T. C. Marsh
K. Aufderheide
W. Ringlien
Introduction
338
1
Indirect Immunofluorescence and Confocal Microscopy
338
6
Fluorescence in Situ Hybridization Detection of Tetrahymena thermophila rDNA for Confocal Microscopy
344
3
Live Fluorescence Microscopy
347
14
References
358
3
Confocal Imaging of Drosophila Embryos
Stephen W. Paddock
Introduction
361
2
Preparation of the Embryos
363
2
Imaging Embryos
365
5
Image Presentation
370
2
Multiple Labels
372
3
Conclusions
375
5
References
376
4
Confocal Fluorescence Microscopy of the Cytoskeleton of Amphibian Oocytes and Embryos
David L. Gard
Introduction
380
3
Fixation of Xenopus Oocytes and Eggs for Confocal Fluorescence Microscopy of the Cytoskeleton
383
7
Processing Xenopus Oocytes and Eggs for ``Whole-Mount'' Confocal Microscopy
390
7
An Introduction to Confocal Microscopy and 3-D Reconstruction of the Cytoskeleton of Xenopus Oocytes
397
11
Conclusions
408
1
Recipes and Reagents
409
1
Cytoskeletal and Nuclear Probes for Confocal Microscopy of Xenopus Oocytes
410
7
References
413
4
Confocal Fluorescence Microscopy Measurements of pH and Calcium in Living Cells
Kay-Pong Yip
Ira Kurtz
Introduction
417
1
Measurement of pHi with BCECF
418
2
Design of a Dual-Excitation Laser Scanning Confocal Fluorescence Microscope for Measuring pHi with BCECF
420
2
Measurement of pHi in the Cortical Collecting Tubule
422
2
Measurement of Intracellular Ca2+ in the Perfused Afferent Arteriole
424
2
Future Development
426
3
References
426
3
Confocal and Nonlinear Optical Imaging of Potentiometric Dyes
Leslie M. Loew
Paul Campagnola
Aaron Lewis
Joseph P. Wuskell
Introduction
429
1
Overview of Fluorescent Methods for Measuring Membrane Potential
430
4
Application of Laser Scanning Microscopy to Quantitative Imaging of Membrane Potential
434
14
Perspectives
448
5
References
449
4
Measurement of Intracellular Ca2+ Concentration
Nicolas Demaurex
Serge Arnaudeau
Michal Opas
Introduction
453
2
Ratio Measurement of pCai with Visible-Light [Ca2+] Indicators
455
5
Measurement of Organellar pCai with Genetically Engineered [Ca2+] Indicators
460
15
References
470
5
Running and Setting Up a Confocal Microscope Core Facility
Susan DeMaggio
Introduction
475
1
Planning Your Facility
476
1
Scheduling
477
1
Training Operators and Users
478
3
Instrument Care
481
1
Data Storage, Analysis, and Handling
482
1
Core Issues
483
2
Ancillary Services
485
1
Resources
485
1
Summary
485
2
Index
487
14
Volumes in Series
501