Fluorescence microscopy [electronic resource] : from principles to biological applications / edited by Ulrich Kubitscheck.
- Weinheim [Germany] : Wiley-Blackwell, 2013.
1 online resource (432 p.)
- Fluorescence microscopy.
- Electronic books.
- This is the first title on the topic designed specifically to allow students and researchers with no background in physics to understand novel light microscopy techniques. Each chapter is written by a renowned expert with many contributions by pioneers in the field, but the editors have ensured that all commonly used methods are covered and that each chapter is comprehensible for non-experts and conforms to the same easily legible standard and style. A companion website with additional examples and video material makes this a valuable teaching resource.
- Fluorescence Microscopy; Contents; Preface; List of Contributors; 1 Introduction to Optics and Photophysics; 1.1 Interference: Light as a Wave; 1.2 Two Effects of Interference: Diffraction and Refraction; 1.3 Optical Elements; 1.3.1 Lenses; 1.3.2 Metallic Mirror; 1.3.3 Dielectric Mirror; 1.3.4 Pinholes; 1.3.5 Filters; 1.3.6 Chromatic Reflectors; 1.4 The Far-Field, Near-Field, and Evanescent Waves; 1.5 Optical Aberrations; 1.6 Physical Background of Fluorescence; 1.7 Photons, Poisson Statistics, and AntiBunching; References; 2 Principles of Light Microscopy; 2.1 Introduction
2.2 Construction of Light Microscopes2.2.1 Components of Light Microscopes; 2.2.2 Imaging Path; 2.2.3 Magnification; 2.2.4 Angular and Numerical Aperture; 2.2.5 Field of View; 2.2.6 Illumination Beam Path; 2.3 Wave Optics and Resolution; 2.3.1 Wave Optical Description of the Imaging Process; 2.3.2 The Airy Function; 2.3.3 Point Spread Function and Optical Transfer Function; 2.3.4 Lateral and Axial Resolution; 2.3.5 Magnification and Resolution; 2.3.6 Depth of Field and Depth of Focus; 2.3.7 Over- and Under Sampling; 2.4 Apertures, Pupils, and Telecentricity; 2.5 Microscope Objectives
2.5.1 Objective Lens Design2.5.2 Light Collection Efficiency and Image Brightness; 2.5.3 Objective Lens Classes; 2.5.4 Immersion Media; 2.5.5 Special Applications; 2.6 Contrast; 2.6.1 Dark Field; 2.6.2 Phase Contrast; 2.6.3 Interference Contrast; 2.6.4 Advanced Topic: Differential Interference Contrast; 2.7 Summary; Acknowledgments; References; 3 Fluorescence Microscopy; 3.1 Features of Fluorescence Microscopy; 3.1.1 Image Contrast; 3.1.2 Specificity of Fluorescence Labeling; 3.1.3 Sensitivity of Detection; 3.2 A Fluorescence Microscope; 3.2.1 Principle of Operation
3.2.2 Sources of Exciting Light3.2.3 Optical Filters in a Fluorescence Microscope; 3.2.4 Electronic Filters; 3.2.5 Photodetectors for Fluorescence Microscopy; 3.2.6 CCD-Charge-Coupled Device; 3.2.7 Intensified CCD (ICCD); 3.2.8 Electron-Multiplying Charge-Coupled Device (EMCCD); 3.2.9 CMOS; 3.2.10 Scientific CMOS (sCMOS); 3.2.11 Features of CCD and CMOS Cameras; 3.2.12 Choosing a Digital Camera for Fluorescence Microscopy; 3.2.13 Photomultiplier Tube (PMT); 3.2.14 Avalanche Photodiode (APD); 3.3 Types of Noise in a Digital Microscopy Image; 3.4 Quantitative Fluorescence Microscopy
3.4.1 Measurements of Fluorescence Intensity and Concentration of the Labeled Target3.4.2 Ratiometric Measurements (Ca++, pH); 3.4.3 Measurements of Dimensions in 3D Fluorescence Microscopy; 3.4.4 Measurements of Exciting Light Intensity; 3.4.5 Technical Tips for Quantitative Fluorescence Microscopy; 3.5 Limitations of Fluorescence Microscopy; 3.5.1 Photobleaching; 3.5.2 Reversible Photobleaching under Oxidizing or Reducing Conditions; 3.5.3 Phototoxicity; 3.5.4 Optical Resolution; 3.5.5 Misrepresentation of Small Objects; 3.6 Current Avenues of Development; References; Further Reading
Recommended Internet Resources
- Description based upon print version of record.
Includes bibliographical references and index.
- Kubitscheck, Ulrich.
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