Handbook of biological confocal microscopy:
Gespeichert in:
| Format: | Tagungsbericht Buch |
|---|---|
| Sprache: | English |
| Veröffentlicht: |
New York [u.a.]
Plenum Press
1990
|
| Ausgabe: | Rev. ed. |
| Schlagworte: | |
| Online-Zugang: | Inhaltsverzeichnis |
| Beschreibung: | Literaturangaben |
| Beschreibung: | XIII, 232 S. Ill., graph. Darst. |
| ISBN: | 0306435381 |
Internformat
MARC
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| 245 | 1 | 0 | |a Handbook of biological confocal microscopy |c ed. by James B. Pawley |
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| 264 | 1 | |a New York [u.a.] |b Plenum Press |c 1990 | |
| 300 | |a XIII, 232 S. |b Ill., graph. Darst. | ||
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| 650 | 4 | |a Microscopie confocale - Congrès | |
| 650 | 4 | |a Confocal microscopy |v Congresses | |
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Datensatz im Suchindex
| _version_ | 1804118730115907584 |
|---|---|
| adam_text | HANDBOOK OF
BIOLOGICAL
CONFOCAL MICROSCOPY
Edited by
James B Pawley
Integrated Microscopy Resource for Biomedical Research
University of Wisconsin-Madison
Madison, Wisconsin
REVISED EDITION
PLENUM PRESS • NEW YORK AND LONDON
Contents
CHAPTER 1: FOUNDATIONS OF CONFOCAL
SCANNED IMAGING IN LIGHT MICROSCOPY 1-14
Shinya Inoue
Light microscopy 1
Lateral resolution 1
Axial resolution 3
Depth of field 3
Confocal imaging 4
Impact of video 5
The Nipkow disk 5
Electron-beam scanning TV 5
Impact of modern video 6
Lasers and microscopy 6
Holography 6
Laser illumination 1
Laser-illuminated confocal microscopes 9
Laser scanning confocal microscope 9
Is laser scanning confocal microscopy a cure-all? 10
Speed of image or data acquisition 10
Depth of field in phase dependent imaging 11
Some other optical and mechanical factors
affecting confocal microscopy 11
Lens aberration 11
Unintentional beam deviation 12
Note added in proof 12
Acknowledgment 13
References 13
CHAPTER 2: FUNDAMENTAL LIMITS IN
CONFOCAL MICROSCOPY 15-26
James Pawley ~—
Introduction 1S
What limits? 15
Counting statistics 16
Source brightness 16
Specimen response 16
A typical problem 16
Practical photon efficiency 17
Losses in the optical system 18
Objectives 18
Mirrors 18
Pinhole 19
Is the confocal pinhole a good thing ? 19
Features of the confocal pinhole 19
Detection and measurement losses 20
The detector 20
The PMT 20
Solid-state photon detectors 21
Digitization 21
Evaluating photon efficiency 21
Resolution: how much is enough? 22
Can resolution be too high? 22
Limitations imposed by spatial and temporal
quantization 23
Aliasing 24
Pixel shape 24
Blind spots 24
Practical considerations relating resolution to
distortion
Summary
Acknowledgements
References
27-39
CHAPTER 3: QUANTITATIVE FLUORESCENCE
IMAGING WITH LASER SCANNING CONFOCAL
MICROSCOPY
K Sam Wells, David R Sandison, James Stricter and
Watt W Webb
The promise of scanning confocal fluorescence
microscopy 27
Optical transfer efficiency 28
Methods of measurement of optical transfer
efficiencies 29
Confocal spatial filtering for depth of field
compromises optical transfer efficiency 30
Optical aberrations in fluorescence LSCM 31
Chromatic aberrations in fluorescence LSCM
measurements 32
Photodynamic effects 33
Theory 34
Population rate equations 34
Experiment 35
Fluorescence photobleaching recovery with LSCM 37
Conclusion 38
Acknowledgments 39
References 39
CHAPTER 4: THE PIXELATED IMAGE 41-51
Robert H Webb and C Kathleen Dorey
Introduction 41
Pixelation 41
Optical resolution: the resel 41
Pixel 42
Gray level 42
In between 42
Matching image spatial characteristics 42
The Nyquist theorem 42
Hyper-resolution: oversampling 43
The resel/pixel ratio 44
Diagonal dropout 44
Pixel shape distortion 45
Aliasing 45
The mechanics of pixelation 46
Matching image intensity characteristics 49
Gray scale 49
Detection 49
Display 49
Color displays 50
Caveats 50
Spectral variation of detectors 50
Automatic gain control 50
Strategy for magnification and resolution 50
Acknowledgements 50
References 50
IX
X Contents
CHAPTER 5: LASER SOURCES FOR CONFOCAL
MICROSCOPY 53-67
Enrico Grat ton and Martin J vandeVen
Introduction 53
Laser power requirements 53
The basic laser 54
Principle of operation 54
Laser modes: longitudinal and transversal 54
Polarization 55
Coherent properties of laser light 55
Temporal coherence 55
Coherence length 55
Spatial coherence 55
Coherence surface 55
Coherence volume 55
Pumping power requirements 55
Heat removal 56
Other installation requirements 56
Types of lasers 56
Continuous wave (cw) lasers 56
Gas lasers 58
Argon-ion 58
Krypton 59
Helium-neon 59
Helium-cadmium 59
Dye lasers 60
Solid state lasers 60
Semiconductor or diode injection lasers 60
Diode-pumped lasers 60
Tunable solid state laser 61
Pulsed lasers 61
Nitrogen lasers 61
Excimer lasers 61
Metal vapor lasers 61
Q-switched lasers 61
Trends in time-resolved spectroscopy applied to
microscopy 63
Wavelength expansion through non-linear
techniques 63
Spatial beam characteristics 63
Intensity fluctuations of cw lasers - 64
Maintenance 64
Maintenance of active laser media 64
Laser tubes 64
Gases 64
Dyes 64
Laser rods 65
Maintenance of pumping media 65
Maintenance of the optical resonator 65
Maintenance of other system components 65
Cooling water 65
External optics 65
Safety precautions 65
Curtains 65
Screens 65
Beam stops 66
Acknowledgement 66
References 66
CHAPTER 6: NON-LASER ILLUMINATION FOR
CONFOCAL MICROSCOPY 69-76
Victor Chen
Introduction 69
Why use non-laser sources? 69
Wavelength 69
Coherence 70
Which types of confocal microscope can use non
laser sources?
Characteristics of non-laser light sources
Wavelengths available
Source radiance
Source stability
Source coherence
Source distribution
Collecting the light and relaying it to specimen
Illumination of the specimen: a basic part of
microscopy
Tandem scanning: basic description
Single-sided disk scanning: basic description
How do you uniformly illuminate both the
objective back focal plane and the inter
mediate image plane?
Scrambling and filtering the light
Measuring what comes through the illumination
system
Exposure time and source brightness
Stationary specimens
What if the specimen is moving or changing?
Incoherent laser light sources for confocal
microscopy
References 76
CHAPTER 7: OBJECTIVE LENSES FOR
CONFOCAL MICROSCOPY 77-86
H Ernst Keller
Abstract 77
Aberrations of refractive systems 77
Defocusing 78
Monochromatic aberrations 79
Spherical aberrations 79-
Coma 80
Astigmatism 80
Flatness of field 80
Distortion 81
Chromatic aberrations 82
Longitudinal chromatic aberration 82
Lateral chromatic aberration (LCA) or
chromatic magnification difference 83
Finite versus infinity optics 84
Optical materials 85
Anti-reflection coatings 85
Conclusion 86
CHAPTER 8: SIZE AND SHAPE OF THE
CONFOCAL SPOT: CONTROL AND RELATION
TO 3D IMAGING AND IMAGE PROCESSING
G J Brakenhoff, K Visscher and HTM van der Voort
Abstract
Introduction
Pinholes and optical probe formation
Practical use of variable pinholes
Experimental axial confocal response
Comments and conclusions
References
87-91
CHAPTER 9: THE INTERMEDIATE OPTICAL
SYSTEM OF LASER-SCANNING CONFOCAL
MICROSCOPES
Ernst H K Stelzer
Design principles of confocal systems
Overview
93-103
Contents xi
Microscope objectives 93
Position of the pivot point 93
Position of the detector pinhole 94
Practical requirements 95
Illumination 95
Detection 96
Distortion 96
Evaluation of illumination/detection systems 96
Influence of optical elements on the properties of
light 96
Errors caused by optical elements 96
Evaluation of optical arrangements 97
Evaluation of scanner arrangements 98
Disk scanners 99
Object scanners 100
Attachment to microscopes 100
Merit functions 100
Requirements for multi-fluorescence experiments 101
Special optical elements 101
Multi-mode optical glass fibers 101
Single-mode polarization-preserving glass fibers 101
Polarizing elements 101
Mechanical scanners 102
Acousto-optical scanners 102
Conclusions 102
Acknowledgements 103
References 103
CHAPTER 10: INTERMEDIATE OPTICS IN
NIPKOW DISK MICROSCOPES 105-111
Gordon S Kino
The tandem scanning reflected light microscope
(TSRLM) 105
The real-time scanning optical microscope (RSOM) 106
Images of the eye 106
Pinhole size 106
Pinhole spacing 109
Illumination efficiency and_ reflection from the disk 110
Internal reflections -- 110
Acknowledgements 111
References 111
CHAPTER 11: THE ROLE OF THE PINHOLE IN
CONFOCAL IMAGING SYSTEMS 113-126
Tony Wilson
Introduction 113
The optical sectioning property 113
The optical sectioning property with a finite-sized
circular detector and coherent light 113
Lateral resolution as a function of effective
detector size 117
The role of aberrations 117
Images with a finite-sized detector 118
Extended-focus and auto-focus imaging with a
finite-sized detector 118
Height imaging with a finite-sized pinhole 118
Alternative detector geometries 119
Noise 121
Fluorescence imaging 123
Conclusions 125
References 125
CHAPTER 12: PHOTON DETECTORS FOR
CONFOCAL MICROSCOPY 127-139
Jonathan Art
Introduction 127
The quantal nature of light 127
Interaction of photons with materials 127
Photoconductivity 128
Photovoltaic 128
Charge coupled devices 129
Photoemissive 130
Image dissector 130
Micro channel plate 130
Noise internal to detectors 131
Statistics of photon flux and detectors 132
Representing the pixel value 133
Conversion techniques 134
Assessment of devices 135
Point detection optimization 135
Field detection optimization 136
Detectors present and future 137
References 138
CHAPTER 13: MANIPULATION DISPLAY AND
ANALYSIS OF THREE-DIMENSIONAL
BIOLOGICAL IMAGES 141 -150
Hans Chen, John W Sedat and David A Agard
Introduction 141
Storage of three-dimensional image data 141
Image enhancement 142
Linear filters 142
Median filters 142
Local contrast enhancement 143
Gradient method 143
Processing methods for displaying 3D data 144
Stereo images 144
3D rotations 144
Rotated projections 145
Pixar displays 145
Contour surface representation 145
Graphic system for 3D image display and analysis 145
Details of PRISM s design and implementation 146
The window system 146
Digital movies 147
Choice of display hardware 147
Model building in PRISM 148
Model building 149
Superimposing the model on a background image 150
Future development and discussion 150
Acknowledgements 150
References 150
CHAPTER 14: THREE-DIMENSIONAL IMAGING
ON CONFOCAL AND WIDE-FIELD
MICROSCOPES 151-161
Walter A Carrrington, Kevin E Fogarty, Larry Lifschitz,
Fredric S Fay
Introduction 151
Signal to noise ratio and resolution 151
Signal strength, photo-damage and photo-bleaching 151
Optical transfer function, resolution and noise 152
Three-dimensional image restoration and confocal
microscopy: a comparison 152
xii Contents
Image restoration methodology 1 S3
Requirements 154
Results 154
Multiple detector confocal miscroscopes 156
Conclusions and recommendations 156
Computer graphics 3D visualization 156
Display of 2D slices of 3D data 156
Volume displays 157
Surface model displays 157
3D perception from 2D displays 157
User interaction and analysis 157
Automated image analysis: feature extraction and
computer vision 157
Thresholding 157
Human interaction and partial automation 158
Fully automated analysis 158
Computer hardware considerations 158
Image acquisition 158
Image restoration 159
Image analysis and display 159
Archival storage 159
Magnetic disks 159
Cartridge magnetic tape 159
Reel magnetic tape 159
Optical disk 159
Networking 159
Conclusion 160
References 160
CHAPTER 15: DIRECT RECORDING OF
STEREOSCOPIC PAIRS OBTAINED DIRECTLY
FROM DISK SCANNING CONFOCAL LIGHT
MICROSCOPES 163-168
Alan Boyde
Summary 163
Introduction 163
Use of a confocal microscope to reduce the depth
of field 163
Optical sectioning in the TSRLM __ 164
Direct photographic recording of the stereo-pair 164
Means for stereo imaging of a layer inside a bulk 164
Fixed tilt angle difference: hand-operated device 164
DC micromotor-controlled stage 164
Piezo-electric control of the lens 165
Top or bottom overlap? 165
Stereopairs generated from one through-focus
pass 165
Topographic mapping 165
Color coding without a computer 165
Depth limitation 166
Particle counting 167
Geometric properties of the stereo images 167
Discussion: TSRLM or LSCM? 167
Acknowledgements 168
References 168
CHAPTER 16: FLUOROPHORES FOR CONFOCAL
MICROSCOPY: PHOTOPHYSICS AND
PHOTOCHEMISTRY 169-178
Roger Y Tsien and Alan Waggoner
Introduction 169
Photophysical problems related to high intensity
excitation 169
Singlet state saturation 169
Triplet state saturation 172
Contaminating background signals 172
Rayleigh and Raman scattering 172
Autofluorescence from endogenous fluorophores 172
What is the optimal intensity? 172
Photodestruction of fluorophores and biological
specimens 172
Dependency on intensity or its time integral? 173
Theory 173
Experiment 173
Protective agents 174
Strategies for signal optimization in the face of
photobleaching 174
Light collection efficiency 174
Spatial resolution 175
Fluorophore concentration 175
Choice of fluorophore 175
Fluorescent indicators for dynamic intracellular
parameters 175
Membrane potentials 175
Ion concentrations 176
Wavelength ratioing 176
pH indicators 176
Ca1+ indicators 176
Other forms of ratioing 177
Future developments? 177
Acknowledgments 177
References 177
CHAPTER 17: IMAGE CONTRAST IN CONFOCAL
LIGHT MICROSCOPY 179-195
P C Cheng and R G Summers
Introduction 179
Sources of contrast 179
Confocal microscopy in back scattered mode 180
Signal formation 180
Backscattered light contrast on stained specimens 181
Reflection contrast on non-biological specimens 182
Backscatter contrast on living specimens 182
The effect of overlying structures 182
Absorption contrast 184
Artificial contrast 184
Transmitted confocal image 186
Confocal microscopy in epi-fluorescent mode 189
Countermeasures 193
Acknowledgement 195
References 196
CHAPTER 18: GUIDING PRINCIPLES OF
SPECIMEN PRESERVATION FOR CONFOCAL
FLUORESCENCE MICROSCOPY 197-205
Robert Bacallao, Morgane Bomsel, Ernst H K Stelzer and
Jan De Mey
Introduction 197
Critical evaluation of fixation and mounting
methods 197
Theoretical considerations 197
The use of the cell height to evaluate the fixation
method 198
The use of cell height to evaluate mounting media 199
Well defined structures can be used to evaluate
fixation methods 199
Comparison of in vivo labeled cell organelles with
immunolabeled cell organelles 200
Contents xiii
Fixation methods 201 Optical Sectioning: Experimental and Theoretical 210
Glutaraldehyde fixation 201 Scanning Mirror/Slit System 210
Stock solutions 201 Confocal Pinhole System 211
Preparation of stock solutions 201 Theoretical Comparison of Slit and Pinhole
Fixation protocol 201 Systems 211
The pH shift/paraformaldehyde fixation 201 A Possible Improvement in Pinhole Confocal
Stock solutions 201 Systems 212
Preparation of the stock solutions 201 A Possible Improvement in the Slit Scanning
Fixation protocol 201 System 213
Immunofluorescence staining 202 Examples of Images Obtained with the Divided
Mounting the specimen 202 Aperture Scanning Slit System 213
General notes 202 Summary Comparison of Slit and Pinhole Confocal
Labeling samples with two or more probes 203 Systems 213
Ramifications of techniques to preserve the Slit system 213
specimens Inherent advantages 213
Conclusion 204 Practical advantages of divided aperture system
Acknowledgements 204 as described 214
References 204 Single pinhole confocal systems 214
Inherent advantages 214
References 214
CHAPTER 19: A COMPARISON OF VARIOUS
OPTICAL SECTIONING METHODS: THE
SCANNING SLIT CONFOCAL MICROSCOPE 207-214 BIBLIOGRAPHY ON CONFOCAL MICROSCOPY 215-227
Charles J Koester Robert H Webb
Introduction 207
Non-Confocal Optical Sectioning 207
Confocal Optical Sectioning 208 INDEX 229-231 Scanning Mifror/Slit Microscope 209
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| genre | (DE-588)1071861417 Konferenzschrift 1989 San Antonio Tex. gnd-content |
| genre_facet | Konferenzschrift 1989 San Antonio Tex. |
| id | DE-604.BV004610551 |
| illustrated | Illustrated |
| indexdate | 2024-07-09T16:14:56Z |
| institution | BVB |
| institution_GND | (DE-588)3006658-X |
| isbn | 0306435381 |
| language | English |
| oai_aleph_id | oai:aleph.bib-bvb.de:BVB01-002832443 |
| oclc_num | 21444071 |
| open_access_boolean | |
| owner | DE-12 DE-19 DE-BY-UBM DE-355 DE-BY-UBR DE-20 DE-188 |
| owner_facet | DE-12 DE-19 DE-BY-UBM DE-355 DE-BY-UBR DE-20 DE-188 |
| physical | XIII, 232 S. Ill., graph. Darst. |
| publishDate | 1990 |
| publishDateSearch | 1990 |
| publishDateSort | 1990 |
| publisher | Plenum Press |
| record_format | marc |
| spelling | Handbook of biological confocal microscopy ed. by James B. Pawley Rev. ed. New York [u.a.] Plenum Press 1990 XIII, 232 S. Ill., graph. Darst. txt rdacontent n rdamedia nc rdacarrier Literaturangaben Microscopia larpcal Microscopie confocale - Congrès Confocal microscopy Congresses Microscopy Congresses Biologie (DE-588)4006851-1 gnd rswk-swf Konfokale Mikroskopie (DE-588)4336446-9 gnd rswk-swf Biowissenschaften (DE-588)4129772-6 gnd rswk-swf Mikroskopie (DE-588)4039238-7 gnd rswk-swf (DE-588)1071861417 Konferenzschrift 1989 San Antonio Tex. gnd-content Mikroskopie (DE-588)4039238-7 s Biologie (DE-588)4006851-1 s DE-604 Konfokale Mikroskopie (DE-588)4336446-9 s Biowissenschaften (DE-588)4129772-6 s 1\p DE-604 Pawley, James B. Sonstige oth Confocal Microscopy Workshop 1989 San Antonio, Tex. Sonstige (DE-588)3006658-X oth HEBIS Datenaustausch application/pdf http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=002832443&sequence=000002&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA Inhaltsverzeichnis 1\p cgwrk 20201028 DE-101 https://d-nb.info/provenance/plan#cgwrk |
| spellingShingle | Handbook of biological confocal microscopy Microscopia larpcal Microscopie confocale - Congrès Confocal microscopy Congresses Microscopy Congresses Biologie (DE-588)4006851-1 gnd Konfokale Mikroskopie (DE-588)4336446-9 gnd Biowissenschaften (DE-588)4129772-6 gnd Mikroskopie (DE-588)4039238-7 gnd |
| subject_GND | (DE-588)4006851-1 (DE-588)4336446-9 (DE-588)4129772-6 (DE-588)4039238-7 (DE-588)1071861417 |
| title | Handbook of biological confocal microscopy |
| title_auth | Handbook of biological confocal microscopy |
| title_exact_search | Handbook of biological confocal microscopy |
| title_full | Handbook of biological confocal microscopy ed. by James B. Pawley |
| title_fullStr | Handbook of biological confocal microscopy ed. by James B. Pawley |
| title_full_unstemmed | Handbook of biological confocal microscopy ed. by James B. Pawley |
| title_short | Handbook of biological confocal microscopy |
| title_sort | handbook of biological confocal microscopy |
| topic | Microscopia larpcal Microscopie confocale - Congrès Confocal microscopy Congresses Microscopy Congresses Biologie (DE-588)4006851-1 gnd Konfokale Mikroskopie (DE-588)4336446-9 gnd Biowissenschaften (DE-588)4129772-6 gnd Mikroskopie (DE-588)4039238-7 gnd |
| topic_facet | Microscopia Microscopie confocale - Congrès Confocal microscopy Congresses Microscopy Congresses Biologie Konfokale Mikroskopie Biowissenschaften Mikroskopie Konferenzschrift 1989 San Antonio Tex. |
| url | http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=002832443&sequence=000002&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA |
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