Light Sheet Fluorescence Microscopy -

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Résumé :

Foreword by Ernst H. K. Stelzer xvii

Preface xxi

1 Let There be Light Sheet 1
Pavel Toman??k and Emmanuel G. Reynaud

1.1 Historical Context of Light Sheet Microscopy - Ultramicroscopy 1

1.2 Light Sheet Imaging Across the Twentieth Century 3

1.3 And here Comes the Flood 3

1.4 The Building of a Community 6

References 8

2 Illumination in Light Sheet Fluorescence Microscopy 11
Rory M. Power and Jan Huisken

2.1 Introduction 11

2.2 Axial Resolution and Optical Sectioning in Light Sheet Microscopy 13

2.2.1 The Point Spread Function in Fluorescence Microscopy 13

2.2.2 The Point Spread Function in Light Sheet Fluorescence Microscopy 14

2.3 Light Sheet Dimensions 16

2.3.1 Gaussian Optics Description of Beam Focusing (x,z Axes) 18

2.3.2 Methods of Light Sheet Production (y Axis) 20

2.4 Practical Light Sheet Generation 21

2.4.1 Static and Pivoted Light Sheets 21

2.4.2 Scanned Light Sheets 27

2.5 Degradation of the Light Sheet in Tissue 29

2.5.1 Absorption of the Light Sheet in Tissue 29

2.5.2 Refraction of the Light Sheet in Tissue 31

2.5.3 Scattering of the Light Sheet in Tissue 31

2.6 Challenges and Benefits of Light Sheet Modes 32

2.6.1 Parallelization of the Light Sheet 32

2.6.2 Image Artifacts Arising from Light Sheet Illumination 34

2.6.3 Homogeneity of Light Sheet Illumination 35

2.6.4 Robustness and Simplicity of Light Sheet Generation 39

2.6.5 The Merits of Static, Pivoted, and Scanned Light Sheets 39

2.7 Multiphoton Excitation 40

2.7.1 Two-Photon Light Sheets 40

2.7.2 Two-Photon Light Sheet Dimensions 41

2.7.3 Comparison with One-Photon Light Sheet Microscopy 45

2.7.4 Comparison with Laser-Scanning Multiphoton Microscopy 47

2.8 Multi-View Illumination 48

2.9 High-Resolution Imaging 51

2.9.1 Geometric Limitations for High-Resolution Imaging 51

2.9.1.1 Oblique Light Sheets 52

2.9.1.2 Reflected Light Sheets 52

2.9.2 Diffractive Limitations for High-Resolution Imaging 54

2.9.2.1 Bessel Beams 54

2.9.2.2 Axially Swept Beams 59

2.9.2.3 Photophysical Approaches 61

2.10 Conclusions 61

References 63

3 A Small Guide on How to Mount a Sample in a Light-Sheet Microscope 67
Francesco Pampaloni, Edward Lachica, Jacques Paysan, and Emmanuel G. Reynaud

3.1 Introduction 67

3.2 A Few Basic Rules 68

3.2.1 Rule 0 - Don't Panic! Become Enthusiastic! 68

3.2.2 Rule 1 - Keep it Clean 68

3.2.3 Rule 2 - The Light Comes Sideways 69

3.2.4 Rule 3 - The Theory does not Apply to your Sample 71

3.2.5 Rule 4 - Sample Geometry Matters 71

3.2.6 Rule 5 - Know Your System Well 72

3.2.7 Rule 6 - How Does Your Sample Move? 73

3.2.8 Rule 7 - What Was Lost? 73

3.2.9 Rule 8 - Consistency is Key 74

3.3 The Light-Matter Conundrum 74

3.4 Hydrogels 75

3.4.1 Preparation 76

3.5 Glues 77

3.6 Sample Holders 77

3.7 Clearing 79

3.8 Cleaning, Labelling, and Storing Samples 83

3.9 An Example: Time-lapse Live Imaging of Three-dimensional Cultures 84

3.9.1 Environmental Control: Temperature, pH, Oxygenation 84

3.9.2 Perfusion-based Environmental Control 88

3.9.3 Sample Holders for the Live Imaging of Three-dimensional Cell Cultures 91

3.9.3.1 Agarose Beakers 91

3.9.3.2 FEP-foil Sample Holders 91

3.9.4 References 93

3.10 A Bit of Literature 94

3.10.1 Reference Guides 95

3.10.2 Your Favorite Models 95

3.10.3 Others 9...

Biographie:

Emmanuel G. Reynaud, PhD, is Assistant Professor in Bioengineering in Biology at University College Dublin, Ireland.

Pavel Toman??k, PhD, is Senior Permanent Research Group Leader at the Max Planck Institute of Molecular Cell Biology and Genetics in Dresden, Germany, and Director of the CEITEC Brno, Czechia. ...

Sommaire:
Light Sheet Fluorescence Microscopy

An indispensable guide to a novel, revolutionary fluorescence microscopy technique!

Light sheet fluorescence microscopy has revolutionized microscopy, since it allows scientists to perform experiments in an entirely different manner and to record data that had not been accessible before. With contributions from noted experts in the fields of physics, biology, and computer science, Light Sheet Fluorescence Microscopy is a unique guide that offers a practical approach to the subject, including information on the basics of light sheet fluorescence microscopy, instrumentation, applications, sample preparation, and data analysis.

Comprehensive in scope, the book is filled with the cutting-edge methods as well as valuable insider tips. Grounded in real-world applications, the book includes chapters from major manufacturers that explores their recent systems and developments. In addition, the book hightlights a discussion of a do-it-yourself light sheet microscope, making the technique affordable for every laboratory.

This important textbook:

• Serves as an easy-to-understand introduction to light sheet-based fluorescence
• Includes numerous tips and tricks for advanced practitioners
• Provides in-depth information on hardware and software solutions for a straightforward implementation of light sheet fluorescence microscopy in the lab
• Includes chapters from the major manufacturers including Zeiss, Leica, Lavision Biotech, Phase View, and Asi

Aimed at cell biologists, biophysicists, developmental biologists, and neuro-biologists, Light Sheet Fluorescence Microscopy offers a comprehensive overview of the most recent applications of this microscopy technique....