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E-grāmata: Practical Guide to Optical Microscopy

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(Durham University, UK)
  • Formāts: 278 pages
  • Izdošanas datums: 14-Jun-2019
  • Izdevniecība: CRC Press
  • Valoda: eng
  • ISBN-13: 9781351630368
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Practical Guide to Optical MicroscopyPalielināt
  • Formāts: 278 pages
  • Izdošanas datums: 14-Jun-2019
  • Izdevniecība: CRC Press
  • Valoda: eng
  • ISBN-13: 9781351630368
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Choice Recommended Title, March 2020

Optical microscopy is used in a vast range of applications ranging from materials engineering to in vivo observations and clinical diagnosis, and thanks to the latest advances in technology, there has been a rapid growth in the number of methods available.

This book is aimed at providing users with a practical guide to help them select, and then use, the most suitable method for their application. It explores the principles behind the different forms of optical microscopy, without the use of complex maths, to provide an understanding to help the reader utilise a specific method and then interpret the results. Detailed physics is provided in boxed sections, which can be bypassed by the non-specialist.

It is an invaluable tool for use within research groups and laboratories in the life and physical sciences, acting as a first source for practical information to guide less experienced users (or those new to a particular methodology) on the range of techniques available.

Features:





The first book to cover all current optical microscopy methods for practical applications Written to be understood by a non-optical expert with inserts to provide the physical science background Brings together conventional widefield and confocal microscopy, with advanced non-linear and super resolution methods, in one book

To learn more about the author please visit here.

Recenzijas

"Although one might assume that many books on microscopy already existed, this reviewer was pleasantly surprised to discover here just how far and how quickly the field has recently moved. Girkin (Durham Univ.) provides a historical overview in the early chapters; though brief, it will be a helpful refresher or introduction for readers new to the subject. The majority of chapters cover advanced topics, including, for example, light sheet microscopy, multiphoton fluorescence microscopy, and digital holographic microscopy. Many of these techniques are new since the development of modern laser systems. It is unlikely that any single lab will use all the covered methods simultaneously, but this book provides a handy comprehensive reference for a general understanding of how the optical method works in the context of various applications and how the data are collected and analyzed in different scenariosThe book will be broadly useful as an authoritative source, especially as most methods introduced are accompanied by a separate explanation of the underlying physics, evidently to avoid overwhelming readers looking for a more general understanding.

Summing Up: Recommended. Upper-division undergraduates through faculty. Students enrolled in two-year technical programs."

T. P. Owen Jr., Connecticut College in CHOICE, March 2020

Preface xiii
Acknowledgements xv
About the Author xvii
Chapter 1 Introduction
1(12)
1.1 A Historical Perspective
2(4)
1.2 Initial Considerations on Which Method to Choose For a Specific Application
6(4)
1.3 How to Use This Book
10(3)
References
10(3)
Chapter 2 Understanding Light in Optical Microscopy
13(26)
2.1 Introduction to the Physics of Optical Microscopy
13(4)
2.1.1 Light
13(2)
2.1.2 Waves, Wavelength, Frequency and Particles
15(1)
2.1.3 Refractive Index
15(1)
2.1.4 Polarization
16(1)
2.2 The Optics of Microscopy
17(4)
2.2.1 Refraction
17(2)
2.2.2 Interference and Diffraction
19(2)
2.3 Limitations in Optical Microscopy
21(3)
2.3.1 Chromatic Aberration
22(1)
2.3.2 Spherical Aberration
22(2)
2.3.3 Astigmatism
24(1)
2.3.4 Field Curvature
24(1)
2.4 Contrast Mechanisms
24(4)
2.4.1 Absorption
25(1)
2.4.2 Scattering
25(2)
2.4.3 Phase or Refractive Index Changes
27(1)
2.4.4 Fluorescence
27(1)
2.4.5 Fluorescence Lifetime
28(1)
2.4.6 Non-linear Excitation; Harmonic Generation, Raman Scattering
28(1)
2.5 A Brief Introduction to Light Sources for Microscopy
28(2)
2.5.1 Conventional Filament Bulb (Including Metal Halide)
29(1)
2.5.2 Mercury Vapour Bulb
29(1)
2.5.3 Light Emitting Diodes (LEDs)
29(1)
2.5.4 Arc Lamp
29(1)
2.5.5 Lasers
30(1)
2.6 Detection of Light in Microscopy
30(9)
2.6.1 Human Eye and Photographic Film
30(2)
2.6.2 Electronic or Digital Camera
32(2)
2.6.3 Photomultiplier
34(2)
2.6.4 Photodiodes
36(1)
References
36(3)
Chapter 3 Basic Microscope Optics
39(16)
3.1 Introduction
39(1)
3.2 Basic Types of Widefield Optical Microscope
39(3)
3.3 Core Optics of a Widefield Microscope
42(2)
3.4 Optimal Illumination
44(2)
3.5 Microscope Objectives
46(4)
3.6 Image Detection and Recording
50(1)
3.7 Guidelines for Use and Advantages and Disadvantages of a Widefield Microscope
51(4)
Chapter 4 Advanced Widefield Microscopy
55(18)
4.1 Introduction
55(1)
4.2 Polarization Microscopy
55(3)
4.2.1 Practical Implementation of Polarization Microscopy
56(2)
4.2.2 Applications of Polarization Microscopy
58(1)
4.3 Phase Contrast Microscopy
58(4)
4.3.1 Practical Implementation of Phase Contrast Microscopy
59(2)
4.3.2 Applications of Phase Microscopy
61(1)
4.4 Differential Interference Contrast (DIC) Microscopy
62(3)
4.4.1 Practical Implementation of DIC Microscopy
64(1)
4.4.2 Applications of DIC Microscopy
65(1)
4.5 Darkfield Microscopy
65(3)
4.5.1 Practical Implementation of Darkfield Microscopy
66(2)
4.5.2 Practical Applications of Darkfield Microscopy
68(1)
4.6 Fluorescence Microscopy
68(4)
4.6.1 Practical Implementation of Fluorescence Microscopy
68(4)
4.6.2 Practical Applications of Fluorescence Microscopy
72(1)
4.7 Summary and Method Selection
72(1)
Chapter 5 Confocal Microscopy
73(24)
5.1 Principles of Confocal Microscopy
74(1)
5.2 Beam Scanned Confocal System
75(4)
5.3 Filter Selection for Beam Scanned Confocal Systems
79(1)
5.4 Detector Selection for Beam Scanned Confocal Systems
80(2)
5.5 Nipkow or Spinning Disk Confocal Systems
82(1)
5.6 Practical Guidelines to Maximize the Performance of a Confocal Microscope
83(12)
5.6.1 Microscope Choice, Sample Mounting and Preparation
85(2)
5.6.2 Lens Selection
87(1)
5.6.3 Initial Image Capture
88(2)
5.6.4 Optimization
90(3)
5.6.5 Saving Data
93(1)
5.6.6 Routine Maintenance
94(1)
5.7 Reconstruction
95(2)
References
95(2)
Chapter 6 Fluorescence Lifetime Imaging Microscopy (FLIM)
97(20)
6.1 Introduction to Fluorescence Lifetime
97(4)
6.1.1 Absorption and Emission
97(3)
6.1.2 Fluorescence Lifetime
100(1)
6.2 Measurement Techniques
101(8)
6.2.1 Time Correlated Single Photon Counting (TCSPC)
101(2)
6.2.2 Time Gating Electronics
103(1)
6.2.3 Time Gated Camera
104(2)
6.2.4 Phase Measurement
106(1)
6.2.5 Slow Detector Method
107(2)
6.3 Methods of Analysis
109(2)
6.4 Experimental Considerations and Guidelines for Use
111(6)
6.4.1 FLIM for Enhancing Contrast
112(1)
6.4.2 FLIM for FRET and Observing Changes in Lifetime
112(2)
6.4.3 FLIM for Absolute Lifetime Measurement
114(1)
6.4.4 Practical Considerations
115(1)
References
116(1)
Chapter 7 Light Sheet or Selective Plane Microscopy
117(22)
7.1 Introduction
117(2)
7.2 Brief History of Light Sheet Microscopy
119(1)
7.3 Optical Principles
120(3)
7.4 Practical Systems
123(4)
7.4.1 Optical Details
123(2)
7.4.2 Basic Alignment
125(1)
7.4.3 Basic Variations in SPIM
126(1)
7.5 Practical Operation
127(6)
7.5.1 Sample Mounting
127(2)
7.5.2 Basic Operation
129(1)
7.5.3 Correcting Common Faults
130(1)
7.5.4 High Speed Imaging and Synchronization
131(1)
7.5.5 High Throughput SPIM
132(1)
7.6 Spim Imaging Processing
133(1)
7.7 Advanced Spim Methods
134(1)
7.8 What is Spim Good for and Limitations
135(4)
References
136(3)
Chapter 8 Multiphoton Fluorescence Microscopy
139(26)
8.1 Introduction to Multiphoton Excitation
140(5)
8.1.1 Requirements on Light Sources for Multiphoton Excitation
142(1)
8.1.2 Requirements on Photon Detection for Multiphoton Microscopy
143(2)
8.2 Practical Multiphoton Microscopy
145(9)
8.2.1 Wavelength
145(1)
8.2.2 Pulse Width and Dispersion Compensation
146(5)
8.2.3 Average Power
151(1)
8.2.4 Objective Lens Selection
152(1)
8.2.5 Detection
153(1)
8.3 Going from Confocal to Multiphoton Microscopy
154(3)
8.4 Advanced Multiphoton Microscopy
157(8)
8.4.1 Endoscopic Multiphoton Microscopy
157(2)
8.4.2 Adaptive Optics for Aberration Correction
159(1)
8.4.3 Measurement of Two Photon "Dose"
160(2)
References
162(3)
Chapter 9 Harmonic Microscopy
165(10)
9.1 Physical Basis for Harmonic Generation
166(2)
9.2 Practical Harmonic Microscopy
168(3)
9.3 Applications of Harmonic Microscopy
171(4)
References
173(2)
Chapter 10 Raman Microscopy
175(18)
10.1 Physical Basis of the Raman Effect
176(3)
10.2 Coherent Anti-Stokes Raman Scattering (CARS)
179(2)
10.3 Stimulated Raman Scattering (SRS) Microscopy
181(1)
10.4 Practical Raman Microscopy Instrumentation
181(4)
10.5 Practical Cars Microscopy Instrumentation
185(1)
10.6 Techniques and Applications in Raman Microscopy
186(3)
10.7 Techniques and Applications in Cars and Srs Microscopy
189(2)
10.8 When to Consider the Use of Raman Microscopy
191(2)
References
191(2)
Chapter 11 Digital Holographic Microscopy
193(14)
11.1 Physical Basis of the Method
194(3)
11.2 Practical Implementation
197(4)
11.2.1 The Light Source for Holographic Microscopy
198(1)
11.2.2 The Detector for Holographic Microscopy
199(1)
11.2.3 The Reconstruction Algorithm for Holographic Microscopy
200(1)
11.3 Practical Applications of Digital Holographic Microscopy
201(4)
11.3.1 Surface Microscopy
202(1)
11.3.2 Particle Tracking
202(1)
11.3.3 Cell Imaging
203(1)
11.3.4 Total Internal Reflection Digital Holographic Microscopy
203(2)
11.4 Summary
205(2)
References
205(2)
Chapter 12 Super Resolution Microscopy
207(18)
12.1 Introduction
207(1)
12.2 Total Internal Reflection Microscopy
208(4)
12.2.1 Principles of Total Internal Reflection Microscopy
208(3)
12.2.2 Practical Implementations of Total Internal Reflection Microscopy
211(1)
12.2.3 Practical Considerations for Total Internal Reflection Microscopy
211(1)
12.3 Structured Illumination Microscopy (SIM)
212(5)
12.3.1 Principles of Structured Illumination Microscopy
213(1)
12.3.2 Practical Implementations of Structured Illumination Microscopy
214(1)
12.3.3 Practical Considerations for Structured Illumination Microscopy
215(2)
12.4 Localization Microscopy (Storm/Palm)
217(3)
12.4.1 Principles of Localization Microscopy
217(2)
12.4.2 Practical Implementations of Localization Microscopy
219(1)
12.4.3 Practical Considerations for Localization Microscopy
219(1)
12.5 Stimulated Emission and Depletion (STED) Microscopy
220(2)
12.5.1 Principles of STED
220(1)
12.5.2 Practical Implementations of STED
221(1)
12.5.3 Practical Considerations for STED
222(1)
12.6 Selection of Super-Resolution Methods
222(3)
References
223(2)
Chapter 13 How to Obtain the Most from Your Data
225(18)
13.1 Introduction
225(1)
13.2 Basics of Data Collection
226(1)
13.3 Software Considerations
227(4)
13.3.1 Open Source Image Processing Packages
228(1)
13.3.2 Programming Languages for Image Processing
228(1)
13.3.3 Commercial Image Processing Packages
229(2)
13.4 Basics of Data Processing
231(5)
13.4.1 Core Techniques
231(1)
13.4.1.1 Reducing Noise
231(1)
13.4.1.2 Uneven Illumination
232(1)
13.4.1.3 Increasing Contrast
233(2)
13.4.1.4 Enhancing Perceived Detail
235(1)
13.4.1.5 Monochrome Look-Up Tables and the Addition of Colour
236(1)
13.5 Producing Quantified Data from Images
236(4)
13.5.1 Intensity-Based Quantification
238(1)
13.5.2 Spatial-Based Quantification
238(1)
13.5.3 Temporal-Based Quantification
239(1)
13.6 Deconvolution
240(2)
13.7 Summary
242(1)
References
242(1)
Chapter 14 Selection Criteria for Optical Microscopy
243(12)
14.1 Introduction
243(1)
14.2 Basic Selection Guidelines
244(4)
14.3 Specialized Techniques
248(5)
14.3.1 Fluorescence Recovery after Photobleaching (FRAP)
248(3)
14.3.2 Forster Resonant Energy Transfer (FRET)
251(1)
14.3.3 Opto-genetics, Observation of Cell Ablation and Photo-uncaging
252(1)
14.3.4 Imaging of Plants and Plant Cells
253(1)
14.4 Summary
253(2)
References
254(1)
Glossary 255(4)
Index 259
John Girkin is professor of Biophysics at Durham University and Director of the Biophysical Sciences Institute in Durham. He moved to Durham in 2009 to take up this role having previously founded the Centre for Biophotonics at Strathclyde University, Glasgow where he was one of the first leaders at the Institute of Photonics. Originally trained as a physicist at Oxford and with a PhD from Southampton University (in Laser Spectroscopy of Atomic Hydrogen) he worked for ten years in industry including developing the worlds first diode laser retinal photocoagulator and diode pumped Nd:Yag laser. He moved back into academia in 1996 as one of the original Research Team Leaders at the Institute of Photonics, Strathclyde University.

His research focuses on the development of novel optical instrumentation to help solve challenges within the life sciences. His research covers a very broad range of activities from dental imaging in the near infrared through to rapid-genomic screening but an ongoing theme has been in developing and applying optical microscopy methods to life science challenges. His initial focus was in non-linear microscopy and he was one of the first pioneers in the use of adaptive optics in microscopy and has more recently been developing advanced forms of single plane illumination microscope for in vivo zebra fish imaging. A constant within his research has been to select the most suitable method for a specific task.

He has published over 100 peer-reviewed publications, is a Fellow of both the Institute of Physics and the Optical Society of America serving on both national and international review panels in the area of biophotonics and microscopy.
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