Understanding Light Microscopy

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.

About the Author

Acknowledgements

Glossary and definitions

About the Companion Website

Notes

Introduction

Chapter 1  Our eyes and the microscope

Chapter 2  Light

Chapter 3  Basic microscope optics

Chapter 4  Microscope design

Chapter 5  Ergonomics

Chapter 6  Optical aberrations of the microscope

Chapter 7  The microscope objective

Chapter 8  Condensers & eyepieces

Chapter 9  Illumination in the microscope

Chapter 10  Diffraction and image formation in microscopy

Chapter 11  Contrast generation & enhancement

Chapter 12  Reflected–light microscopy

Chapter 13  Polarised–light microscopy: part 1 – theory

Chapter 14  Polarised–light microscopy: part 2 – applied

Chapter 15  Fluorescence microscopy

Chapter 16  Fluorophores & fluorescent proteins

Chapter 17 Optical sectioning & confocal microscopy

Chapter 18  Operating the confocal microscope

Chapter 19 Light–sheet microscopy

Chapter 20  Bleed–through & Spectral unmixing

Chapter 21  Deconvolution

Chapter 22  Multi–photon microscopy

Chapter 23  Total Internal Reflection Fluorescence microscopy

Chapter 24  FRAP and FRET

Chapter 25  Colocalisation

Chapter 26 Super–resolution fluorescence microscopy

Chapter 27 – Choosing a microscope platform and Core Imaging Facilities

Chapter 28  Biological Specimen Preparation

Chapter 29  Materials Specimen Preparation

Chapter 30  Recording the image: part 1 – theory

Chapter 31  Recording the image: part 2 – applied

Appendices

1. Buying and tendering for a light microscope

2. Trouble–shooting poor image quality

3. The Michel–Lévy Interference chart

4. Cleaning and Maintenance

5. Microscope Manufacturer contacts

6. Brief historical notes

7. Timeline

Index

Jeremy Sanderson is a Bio–Imaging specialist at the MRC Harwell Institute Mammalian Genetics Unit in the UK. He is a Fellow of the Royal Microscopical Society and has served both on the Executive Council and the LM Section Committee.