Introduces readers to the enlightening world of the modern light microscope
There have been rapid advances in science and technology over the last decade, and the light microscope, together with the information that it gives about the image, has changed too. Yet the fundamental principles of setting up and using a microscope rests upon unchanging physical principles that have been understood for years. This informative, practical, full-colour guide fills the gap between specialised edited texts on detailed research topics, and introductory books, which concentrate on an optical approach to the light microscope. It also provides comprehensive coverage of confocal microscopy, which has revolutionised light microscopy over the last few decades.
Written to help the reader understand, set up, and use the often very expensive and complex modern research light microscope properly, Understanding Light Microscopy keeps mathematical formulae to a minimum - containing and explaining them within boxes in the text. Chapters provide in-depth coverage of basic microscope optics and design; ergonomics; illumination; diffraction and image formation; reflected-light, polarised-light, and fluorescence microscopy; deconvolution; TIRF microscopy; FRAP & FRET; super-resolution techniques; biological and materials specimen preparation; and more.
- Gives a didactic introduction to the light microscope
- Encourages readers to use advanced fluorescence and confocal microscopes within a research institute or core microscopy facility
- Features full-colour illustrations and workable practical protocols
Understanding Light Microscopy is intended for any scientist who wishes to understand and use a modern light microscope. It is also ideal as supporting material for a formal taught course, or for individual students to learn the key aspects of light microscopy through their own study.
Table of Contents
About the Author ix
Acknowledgements xi
Look-Up Guide to Feature Boxes by Theme xii
Glossary and Definitions xiv
Notes xxiv
Introduction xxvii
1 Our Eyes and the Microscope 1
2 Light 29
3 Basic Microscope Optics 55
4 Microscope Anatomy and Design 75
5 Ergonomics 91
6 Optical Aberrations of the Microscope 101
7 The Microscope Objective 127
8 Condensers and Eyepieces 161
9 Illumination in the Microscope 177
10 Diffraction and Image Formation in Microscopy 211
11 Contrast Generation and Enhancement 243
12 Reflected-Light Microscopy 289
13 Polarised-Light Microscopy: Part 1 - Theory 317
14 Polarised-Light Microscopy: Part 2 - Applied 347
15 Fluorescence Microscopy 383
16 Fluorophores and Fluorescent Proteins 405
17 Optical Sectioning and Confocal Microscopy 425
18 Operating the Confocal Microscope 447
19 Light-Sheet Microscopy 483
20 Bleed-Through and Spectral Unmixing 507
21 Deconvolution 523
22 Multi-Photon Microscopy 543
23 Total Internal Reflection Fluorescence Microscopy 561
24 FRAP and FRET 569
25 Colocalisation 587
26 Super-Resolution Fluorescence Microscopy 613
27 Choosing a Microscope Platform and Core Imaging Facilities 637
28 Biological Specimen Preparation 663
29 Materials Specimen Preparation 687
30 Recording the Image: Part 1 - Theory 707
31 Recording the Image: Part 2 - Applied 733
Appendices
1 Buying, and Tendering for, a Light Microscope 769
2 Troubleshooting Poor Image Quality 773
3 The Michel-Lévy Interference Colour Chart 775
4 Cleaning and Maintenance of the Light Microscope 779
5 Selected Suppliers 783
6 Historical Background 787
7 Timeline of Key Events 791
Index 799
