Fundamentals of light microscopy and electronic imaging:
Gespeichert in:
| Hauptverfasser: | , |
|---|---|
| Format: | Buch |
| Sprache: | English |
| Veröffentlicht: |
Hoboken, NJ
Wiley-Blackwell
2013
|
| Ausgabe: | 2. ed. |
| Schlagworte: | |
| Online-Zugang: | Inhaltsverzeichnis |
| Beschreibung: | XIII, 538 S. Ill., graph. Darst. |
| ISBN: | 9780471692140 |
Internformat
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| 100 | 1 | |a Murphy, Douglas B. |d 1945- |e Verfasser |0 (DE-588)1236293657 |4 aut | |
| 245 | 1 | 0 | |a Fundamentals of light microscopy and electronic imaging |c Douglas B. Murphy ; Michael W. Davidson |
| 250 | |a 2. ed. | ||
| 264 | 1 | |a Hoboken, NJ |b Wiley-Blackwell |c 2013 | |
| 300 | |a XIII, 538 S. |b Ill., graph. Darst. | ||
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Datensatz im Suchindex
| _version_ | 1804148547325526016 |
|---|---|
| adam_text | CONTENTS
Preface
xi
Acknowledgments
xii
1.
FUNDAMENTALS OF LIGHT MICROSCOPY
1
Overview
1
Optical Components of the Light Microscope
1
Aperture and Image Planes in a Focused, Adjusted Microscope
5
Note: Objectives, Eyepieces, and Eyepiece Telescopes
6
Koehler Illumination
9
Adjusting the Microscope for Koehler Illumination
9
Note: Summary of Steps for Koehler Illumination
11
Note: Focusing Oil Immersion Objectives
14
Fixed Tube Length versus Infinity Optical Systems
15
Precautions for Handling Optical Equipment
16
Care and Maintenance of the Microscope
17
Exercise: Calibration of Magnification
17
2.
LIGHT AND COLOR
21
Overview
21
light as a Probe of Matter
21
The Dual Particle- and Wave-like Nature of light
25
The Quality of light
26
Properties of light Perceived by the Eye
27
Physical Basis for Visual Perception and Color
28
Addition and Subtraction Colors
30
Exercise: Complementary Colors
32
VÍ
CONTENTS
3. ILLUMINATORS, FILTERS, AND THE ISOLATION
OF
SPECIFIC WAVELENGTHS
35
Overview
35
Illuminators and Their Spectra
35
Illuminator Alignment and Bulb Replacement
41
Demonstration: Spectra of Common Light Sources
41
Demonstration: Aligning a 100-W Mercury Arc Lamp in an Epi-Illuminator
43
Filters for Adjusting the Intensity and Wavelength of Illumination
45
Effects of Light on Living Cells
50
4.
LENSES AND GEOMETRICAL OPTICS
53
Overview
53
Reflection and Refraction of Light
53
Image Formation by a Simple Lens
56
Note: Real and Virtual Images
57
Rules of Ray Tracing for a Simple Lens
58
Object-Image Math
58
The Principal Aberrations of Lenses
62
Designs and Specifications of Objectives
65
Condensers
71
Oculars
72
Microscope Slides and
Coverslips 73
The Care and Cleaning of Optics
73
Exercise: Constructing and Testing an Optical Bench Microscope
76
5.
DIFFRACTION AND INTERFERENCE IN IMAGE
FORMATION
79
Overview
79
Diffraction and Interference
80
The Diffraction Image of a Point Source of Light
83
The Constancy of Optical Path Length between Object and Image
85
Demonstration: Viewing the Airy Disk with a Pinhole Aperture
85
Effect of Aperture Angle on Diffraction Spot Size
87
Diffraction by a Grating and Calculation of Its Line Spacing,
D
89
Demonstration: The Diffraction Grating
93
Abbé s
Theory for Image Formation in the Microscope
94
A Diffraction Pattern Is Formed in the Rear Aperture of the Objective
97
Demonstration: Observing the Diffraction Image in the Rear Focal Plane of
a Lens
98
Preservation of Coherence: Essential Requirement for Image
Formation
99
Exercise.· Diffraction by Microscope Specimens
101
6.
DIFFRACTION AND SPATIAL RESOLUTION
103
Overview
103
Numerical Aperture
103
CONTENTS
VU
Spatial
Resolution 105
Depth of Field and Depth of Focus
109
Optimizing the Microscope Image: A Compromise between Spatial
Resolution and Contrast
109
Exercise: Resolution
ofStríae
in Diatoms
112
7.
PHASE CONTRAST MICROSCOPY AND DARKFIELD
MICROSCOPY
115
Overview
115
Phase Contrast Microscopy
115
The Behavior of Waves from Phase Objects in Brightfield Microscopy
119
Exercise: Determination of the Intracellular Concentration of Hemoglobin in
Erythrocytes by Phase Immersion Refractometry
128
Darkfield Microscopy
129
Exercise: Darkfield Microscopy
133
8.
PROPERTIES OF POLARIZED LIGHT
135
Overview
135
The Generation of Polarized Light
135
Demonstration: Producing Polarized Light with a Polaroid Filter
137
Polarization by Reflection and Scattering
139
Vectorial
Analysis of Polarized Light Using a Dichroic Filter
139
Double Refraction in Crystals
142
Demonstration: Double Refraction by
a Calcite
Crystal
144
Kinds of Birefringence
145
Propagation of
О
and
E
Wavefronts in
a Biréfringent
Crystal
146
Birefringence in Biological Specimens
148
Generation of EHiptically Polarized Light by
Biréfringent
Specimens
149
9.
POLARIZATION MICROSCOPY
153
Overview
153
Optics of the Polarizing Microscope
155
Adjusting the Polarizing Microscope
156
Appearance of
Biréfringent
Objects in Polarized light
157
Principles of Action of Retardation Plates and Three Popular
Compensators
158
Demonstration: Making a
λ
-Plate
from a Piece of Cellophane
162
Exercise: Determination of Molecular Organization in Biological Structures
Using a Full Wave Plate Compensator
167
10.
DIFFERENTIAL INTERFERENCE CONTRAST
MICROSCOPY AND MODULATION CONTRAST
MICROSCOPY
173
Overview
173
The DIC Optical System
173
Demonstration: The
Atìion
of a Wollaston Prism in Polarized Light
179
Vili
CONTENTS
Modulation Contrast Microscopy
190
Exercise: DIC Microscopy
194
11.
FLUORESCENCE MICROSCOPY
199
Overview
199
Applications of Fluorescence Microscopy
201
Physical Basis of Fluorescence
202
Properties of Fluorescent Dyes
205
Demonstration: Fluorescence of Chlorophyll and Fluorescein
206
Autofluorescence of Endogenous Molecules
211
Demonstration: Fluorescence of Biological Materials under
UV Light
213
Fluorescent Dyes and Proteins in Fluorescence Microscopy
213
Arrangement of Filters and the Epi-Illuminator in the Fluorescence
Microscope
218
Objectives and Spatial Resolution in Fluorescence Microscopy
224
Causes of High Fluorescence Background
225
The Problem of Bleedthrough with Multiply Stained Specimens
227
Quenching, Blinking, and Photobleaching
228
Examining Fluorescent Molecules in Living Cells
230
12.
FLUORESCENCE IMAGING OF DYNAMIC
MOLECULAR PROCESSES
233
Overview
233
Modes of Dynamic Fluorescence Imaging
234
Förster
Resonance Energy Transfer
236
Applications
244
Fluorescence Recovery after Photobleaching
245
TIRF Microscopy: Excitation by an Evanescent Wave
252
Advanced and Emerging Dynamic Fluoresence Techniques
261
13.
CONFOCAL LASER SCANNING MICROSCOPY
265
Overview
265
The Optical Principle of Confocal Imaging
267
Demonstration: Isolation of Focal Plane Signals with a Confocal
Pinhole
271
Advantages of CLSM over Widefield Fluorescence Systems
273
Criteria Defining Image Quality and the Performance of an Electronic
Imaging System
275
Confocal Adjustments and Their Effects on Imaging
277
Photobleaching
286
General Procedure for Acquiring a Confocal Image
286
Performance Check of a Confocal System
288
Fast (Real-Time) Imaging in Confocal Microscopy
288
Spectral Analysis: A Valuable Enhancement for Confocal Imaging
295
CONTENTS
ЇХ
Optical Sectioning by Structured Illumination
297
Deconvolution Microscopy
298
Exercise: Effect of Confocal Variables on Image Quality
304
14.
TWO-PHOTON EXCITATION FLUORESCENCE
MICROSCOPY
307
Overview
307
The Problem of Photon Scattering in Deep Tissue Imaging
308
Two-Photon Excitation Is a Nonlinear Process
309
Localization of Excitation
314
Why Two-Photon Imaging Works
317
Resolution
318
Equipment
319
Three-Photon Excitation
325
Second Harmonic Generation Microscopy
326
15. SUPERRESOLUTION
IMAGING
331
Overview
331
The RESOLFT Concept
333
Single-Molecule Localization Microscopy
334
Structured Illumination Microscopy
343
Stimulated Emission Depletion (STED) Microscopy:
Superresolution
by
PSF
Engineering
349
16.
IMAGING LIVING CELLS WITH THE MICROSCOPE
357
Overview
357
Labeling Strategies for Live-Cell Imaging
358
Control of Illumination
361
Control of Environmental Conditions
365
Optics, Detectors, and Hardware
372
Evaluating Live-Cell Imaging Results
384
Exercise.· Fluorescence Microscopy of Living Tissue Culture Cells
384
17.
FUNDAMENTALS OF DIGITAL IMAGING
389
Overview
389
The Charge-Coupled Device
(CCD Imager)
390
CCD
Designs
396
Note: Interline
CCD Imagers:
The Design of Choice for
Biomedical
Imaging
398
Back-Thinned Sensors
398
EMCCD Cameras: High Performance Design for Greatest Sensitivity
399
Scientific CMOS: The Next Generation of Scientific Imagers
400
Camera Variables Affecting
CCD
Readout and Image Quality
401
Six Terms Define Imaging Performance
404
Aliasing
409
CONTENTS
Color Cameras
410
Exercise: Evaluating the Performance of a CCD Camera
411
18.
DIGITAL IMAGE PROCESSING
415
Overview
415
Preliminaries: Image Display and Data Types
416
Histogram Adjustment
417
Adjusting Gamma
(γ)
to Create Exponential LUTs
421
Flat-Field Correction
421
Image Processing With Filters
425
Signal-to-Noise Ratio
432
The Use of Color
438
Images as Research Data and Requirements for Scientific Publication
442
Exercise: Flat-Field Correction and Determination ofS/N Ratio
448
Appendix A: Answer Key to Exercises
451
Appendix B: Materials for Demonstrations and
Exercises
455
Appendix C: Sources of Materials for Demonstrations
and Exercises
463
Glossary
465
Microscopy Web Resources
509
Recommended Reading
521
References
523
Index
531
|
| any_adam_object | 1 |
| author | Murphy, Douglas B. 1945- Davidson, Michael W. |
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| dewey-ones | 502 - Miscellany |
| dewey-raw | 502.282 |
| dewey-search | 502.282 |
| dewey-sort | 3502.282 |
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| discipline | Allgemeine Naturwissenschaft Physik Biologie |
| edition | 2. ed. |
| format | Book |
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| indexdate | 2024-07-10T00:08:52Z |
| institution | BVB |
| isbn | 9780471692140 |
| language | English |
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| spelling | Murphy, Douglas B. 1945- Verfasser (DE-588)1236293657 aut Fundamentals of light microscopy and electronic imaging Douglas B. Murphy ; Michael W. Davidson 2. ed. Hoboken, NJ Wiley-Blackwell 2013 XIII, 538 S. Ill., graph. Darst. txt rdacontent n rdamedia nc rdacarrier Microscopy Bildverarbeitung (DE-588)4006684-8 gnd rswk-swf Bildgebendes Verfahren (DE-588)4006617-4 gnd rswk-swf Mikroskopie (DE-588)4039238-7 gnd rswk-swf Bildgebendes Verfahren (DE-588)4006617-4 s DE-604 Mikroskopie (DE-588)4039238-7 s Bildverarbeitung (DE-588)4006684-8 s 1\p DE-604 Davidson, Michael W. Verfasser aut Digitalisierung UB Regensburg application/pdf http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=024531130&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 | Murphy, Douglas B. 1945- Davidson, Michael W. Fundamentals of light microscopy and electronic imaging Microscopy Bildverarbeitung (DE-588)4006684-8 gnd Bildgebendes Verfahren (DE-588)4006617-4 gnd Mikroskopie (DE-588)4039238-7 gnd |
| subject_GND | (DE-588)4006684-8 (DE-588)4006617-4 (DE-588)4039238-7 |
| title | Fundamentals of light microscopy and electronic imaging |
| title_auth | Fundamentals of light microscopy and electronic imaging |
| title_exact_search | Fundamentals of light microscopy and electronic imaging |
| title_full | Fundamentals of light microscopy and electronic imaging Douglas B. Murphy ; Michael W. Davidson |
| title_fullStr | Fundamentals of light microscopy and electronic imaging Douglas B. Murphy ; Michael W. Davidson |
| title_full_unstemmed | Fundamentals of light microscopy and electronic imaging Douglas B. Murphy ; Michael W. Davidson |
| title_short | Fundamentals of light microscopy and electronic imaging |
| title_sort | fundamentals of light microscopy and electronic imaging |
| topic | Microscopy Bildverarbeitung (DE-588)4006684-8 gnd Bildgebendes Verfahren (DE-588)4006617-4 gnd Mikroskopie (DE-588)4039238-7 gnd |
| topic_facet | Microscopy Bildverarbeitung Bildgebendes Verfahren Mikroskopie |
| url | http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=024531130&sequence=000002&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA |
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