| Description |
1 online resource (xvi, 119 pages) : illustrations |
| Series |
Tutorial texts in optical engineering ; v. TT 69 |
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SPIE tutorial texts ; TT69
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| Contents |
Symbols and abbreviations -- Preface -- Chapter 1 Confocal fluorescence microscopy -- 1.1. The principle -- 1.2. The theory. 1.2.1. The point spread function (PSF); 1.2.2. Validity of approximations; 1.2.3. The PSF in 3D; 1.2.4. Image formation; 1.2.5. The confocal PSF; 1.2.6. The optical transfer function (OTF) -- 1.3. What is the resolution?. 1.3.1. Resolution and the OTF; 1.3.2. Magnification -- 1.4. Optical aberrations. 1.4.1. Chromatic aberration; 1.4.2. Refractive index mismatch |
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Chapter 2. Implementation -- 2.1. The microscope objective. 2.1.1. Magnification, NA and immersion; 2.1.2. Tube length; 2.1.3. Cover glass; 2.1.4. Microscope objective types; 2.1.5. Working distance; 2.1.6. Transmission -- 2.2. Scanning. 2.2.1. Specimen scanning; 2.2.2. Laser scanning; 2.2.3. The Nipkow disk; 2.2.4. Bilateral scanning -- 2.3. The detection pinhole -- 2.4. Fluorophores. 2.4.1. Absorption and fluorescence; 2.4.2. Molecular energy levels; 2.4.3. Absorption; 2.4.4. Vibrational and rotational relaxation; 2.4.5. Emission; 2.4.6. Phosphorescence; 2.4.7. Bleaching; 2.4.8. Fluorescent proteins -- 2.5. The laser |
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Chapter 3. Practical limits -- Chapter 4. Digitization -- 4.1. Discrete sampling of continuous signals -- 4.2. Digitization in the spatial domain -- 4.3. Digitization in the intensity domain |
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Chapter 5. Miscellaneous topics -- 5.1. PSF measurement. 5.1.1. Preparation protocol -- 5.2. Restoration/deconvolution. 5.2.1. The principle; 5.2.2. Practical considerations; 5.2.3. Practical example -- 5.3. The commercial instrument. 5.3.1. Excitation; 5.3.2. Objectives; 5.3.3. Dichroics; 5.3.4. Detection pinhole; 5.3.5. Detection; 5.3.6. Speed; 5.3.7. Software -- 5.4. Related 3D fluorescence techniques. 5.4.1. 4[pi]-microscopy; 5.4.2. Two-photon absorption microscopy |
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Appendix A. Elements of optical theory. A.1. Geometrical optics; A.2. Aberration theory -- Appendix B. Formulae, relations, and definitions. B.1. Optical coordinates; B.2. Focal field distribution; B.3. Classical theory versus full diffraction theory; B.4. Confocal PSF -- Appendix C. Fourier theory. C.1. Properties of the Fourier transform -- Bibliography -- Index |
| Summary |
This book provides a comprehensive account of the theory of image formation in a confocal fluorescence microscope as well as a practical guideline to the operation of the instrument, its limitations, and the interpretation of confocal microscopy data. The appendices provide a quick reference to optical theory, microscopy-related formulas and definitions, and Fourier theory |
| Bibliography |
Includes bibliographical references (pages 111-114) and index |
| Subject |
Confocal fluorescence microscopy.
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SCIENCE -- General.
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Confocal fluorescence microscopy
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| Form |
Electronic book
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| Author |
Society of Photo-Optical Instrumentation Engineers.
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| ISBN |
9780819481177 |
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0819481173 |
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