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xv | |
Preface |
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xix | |
Acknowledgments |
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xxi | |
About the Authors |
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xxiii | |
About the Illustrator |
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xxv | |
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1 | (8) |
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1.1 Historical Overview of Light Microscopy |
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1 | (1) |
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1.2 Light Microscopic Modalities |
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1 | (3) |
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1 | (1) |
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2 | (1) |
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1.2.3 Phase-Contrast and Differential Interference Contrast |
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2 | (1) |
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1.2.4 Polarization Microscopy |
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3 | (1) |
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1.2.5 Basic Fluorescence Microscopy |
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3 | (1) |
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1.2.6 Axially Selective Illumination Methods |
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4 | (1) |
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1.2.7 Super-Resolution Methods |
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4 | (1) |
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1.3 Historical Overview of Medical Imaging |
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4 | (2) |
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1.4 Medical Imaging Modalities |
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6 | (3) |
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6 | (1) |
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1.4.2 Conventional Radiography and Computed Tomography |
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6 | (1) |
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1.4.3 Planar and Tomographic Nuclear Imaging |
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7 | (1) |
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1.4.4 Magnetic Resonance Imaging (MRI) |
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7 | (1) |
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Suggested Reading and Additional Resources |
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8 | (1) |
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2 Review of Essential Basics |
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9 | (16) |
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9 | (3) |
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2.2 Interactions of Waves and Matter |
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12 | (5) |
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2.2.1 Qualitative Description |
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12 | (1) |
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12 | (1) |
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12 | (1) |
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2.2.1.3 Elastic Scattering |
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12 | (1) |
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13 | (1) |
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2.2.1.5 Change in Wave Speed |
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13 | (1) |
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2.2.2 Reflection and Refraction at Boundaries |
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13 | (4) |
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2.3 Superposition of Waves |
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17 | (2) |
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2.4 Polarization of EM Radiation |
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19 | (6) |
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2.4.1 Types of Polarization |
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19 | (1) |
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2.4.2 Generating and Manipulating Polarized Light |
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19 | (2) |
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Suggested Reading and Additional Resources |
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21 | (1) |
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21 | (4) |
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25 | (156) |
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3 Introduction to Image Formation by the Optical Microscope |
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27 | (22) |
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27 | (1) |
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3.2 Optical Power, Focal Length, and Focal Plane |
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28 | (2) |
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3.3 Types of Lenses and Aberrations |
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30 | (2) |
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3.3.1 Single-Element (Simple) Lenses |
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30 | (1) |
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3.3.2 Multi-Element (Compound) Lenses |
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30 | (2) |
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32 | (5) |
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3.4.1 Qualitative Analysis |
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32 | (1) |
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3.4.2 Quantitative Analysis |
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33 | (1) |
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33 | (1) |
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34 | (3) |
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3.4.2.3 Angular Magnification |
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37 | (1) |
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3.5 The Rudimentary Compound Microscope |
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37 | (2) |
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3.6 The Research-Grade Optical Microscope |
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39 | (10) |
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3.6.1 Kohler Illumination |
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39 | (1) |
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40 | (4) |
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Suggested Reading and Additional Resources |
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44 | (1) |
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44 | (2) |
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46 | (1) |
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A3.1 Construction of a simple compound microscope |
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46 | (3) |
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4 Wave Theory of Image Formation and Resolution |
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49 | (20) |
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49 | (4) |
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4.1.1 Aperture-Induced Spreading |
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49 | (1) |
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4.1.2 Quantifying Resolution |
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49 | (3) |
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4.1.3 Rayleigh's Resolution Criterion |
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52 | (1) |
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53 | (3) |
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4.3 The Significance of Numerical Aperture |
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56 | (2) |
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4.4 Abbe's Experiments: Manipulating Specimen Diffraction Patterns |
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58 | (3) |
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4.5 The Role of Coherence |
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61 | (1) |
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4.6 Mathematical Analysis of Interference from Two Coherent Sources [ Optional] |
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62 | (7) |
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Suggested Reading and Additional Resources |
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64 | (1) |
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64 | (3) |
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67 | (1) |
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A4.1 Generation of Simple Diffraction Patterns |
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67 | (1) |
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67 | (2) |
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5 Contrast Enhancement in Optical Microscopy |
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69 | (26) |
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5.1 Brightfield Microscopy |
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69 | (1) |
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5.1.1 Amplitude Specimens |
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69 | (1) |
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69 | (1) |
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5.2 Contrast-Enhancing Approaches |
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69 | (20) |
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69 | (1) |
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5.2.2 Darkfield Microscopy |
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70 | (2) |
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5.2.2.1 Mechanism of Contrast Generation |
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72 | (1) |
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5.2.2.2 Experimental Implementation |
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72 | (1) |
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5.2.3 Phase-Contrast Microscopy |
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72 | (1) |
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5.2.3.1 Vector Analysis of Phase Specimens |
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73 | (3) |
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5.2.3.2 Specimen Appearance in Positive Phase Contrast |
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76 | (1) |
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5.2.3.3 Experimental Implementation |
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76 | (1) |
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5.2.4 Polarization Microscopy |
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76 | (1) |
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76 | (3) |
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5.2.4.2 Mechanism of Contrast Generation |
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79 | (3) |
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5.2.4.3 Experimental Implementation |
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82 | (1) |
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5.2.5 Differential Interference Contrast |
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83 | (1) |
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5.2.5.1 Mechanism of Contrast Generation |
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83 | (4) |
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5.2.5.2 Experimental Implementation |
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87 | (2) |
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5.3 Comparison of Approaches |
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89 | (6) |
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Suggested Reading and Additional Resources |
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89 | (1) |
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89 | (4) |
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93 | (1) |
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A5.1 Tour of Brightfield, Darkfield, and Phase-Contrast Microscopies |
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93 | (1) |
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A5.2 Introduction to Polarization Microscopy |
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94 | (1) |
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6 Fluorescence Microscopy |
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95 | (24) |
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6.1 Attributes of Fluorophores and Fluorescence |
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95 | (4) |
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6.1.1 Fluorophore Structure |
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95 | (1) |
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6.1.2 Fluorescence Excitation and Emission Spectra |
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95 | (4) |
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6.2 Fluorescence Labeling Techniques |
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99 | (3) |
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6.2.1 Conventional Staining |
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99 | (1) |
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6.2.1.1 Covalent Labeling |
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99 | (1) |
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6.2.1.2 Immunofluorescence Labeling |
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99 | (1) |
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6.2.1.3 Fluorescent Chemical Reagents |
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99 | (1) |
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6.2.2 Genetically Encoded Fluorescent Reporters |
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100 | (2) |
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6.3 The Widefield Fluorescence Microscope |
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102 | (3) |
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6.3.1 The Epi-Fluorescence Optical Train |
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102 | (1) |
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102 | (1) |
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6.3.3 Imaging Modes for Thin Samples |
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102 | (3) |
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6.3.4 The Blur Problem for "Thick" Samples |
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105 | (1) |
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105 | (4) |
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6.4.1 Key Optical Principles of LSCM |
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106 | (1) |
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6.4.2 Instrumentation for LSCM |
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107 | (1) |
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6.4.3 Key Optical Principles of SDCM |
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107 | (1) |
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6.4.4 Instrumentation for SDCM |
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108 | (1) |
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108 | (1) |
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109 | (3) |
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6.6 Comparison of Confocal Microscopy and Deconvolution |
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112 | (7) |
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Suggested Reading and Additional Resources |
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113 | (1) |
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113 | (3) |
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116 | (1) |
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116 | (3) |
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7 Axially Selective Fluorescence Excitation Techniques |
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119 | (16) |
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7.1 Total Internal Reflection Fluorescence Microscopy (TIRFM) |
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119 | (4) |
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7.1.1 Mechanism of Axial Discrimination |
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119 | (1) |
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7.1.2 Experimental Implementation |
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120 | (3) |
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7.2 Light Sheet Fluorescence Microscopy (LSFM) |
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123 | (3) |
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7.2.1 Mechanism of Axial Discrimination |
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123 | (2) |
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7.2.2 Experimental Implementation |
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125 | (1) |
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7.3 Two-Photon Fluorescence Microscopy |
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126 | (5) |
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7.3.1 Two-Photon Excitation |
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126 | (1) |
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7.3.2 Mechanism of Axial Discrimination |
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127 | (2) |
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7.3.3 Application to Deep Imaging |
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129 | (1) |
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7.3.4 Experimental Implementation |
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130 | (1) |
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7.4 Comparison of Approaches |
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131 | (4) |
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Suggested Reading and Additional Resources |
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131 | (1) |
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131 | (4) |
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8 Super-Resolution Fluorescence Techniques |
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135 | (18) |
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8.1 Super-Resolution Structured Illumination Microscopy (SR-SIM) |
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135 | (4) |
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8.1.1 Mechanism of Resolution Enhancement |
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135 | (2) |
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8.1.2 Experimental Implementation |
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137 | (2) |
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8.2 Sequential Readout-Based Approaches |
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139 | (8) |
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8.2.1 Stimulated Emission Depletion (STED) |
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140 | (1) |
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8.2.1.1 Mechanism of Resolution Enhancement |
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140 | (1) |
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8.2.1.2 Spot Engineering via Stimulated Emission |
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141 | (1) |
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8.2.1.3 Experimental Implementation |
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142 | (1) |
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8.2.2 Photoactivated Localization Microscopy (PALM) |
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143 | (1) |
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8.2.2.1 Mechanism of Resolution Enhancement |
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143 | (1) |
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143 | (2) |
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8.2.2.3 Image Resolution and the Nyquist Sampling Criterion |
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145 | (1) |
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8.2.2.4 Experimental Implementation |
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146 | (1) |
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8.3 Comparison of Approaches |
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147 | (6) |
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Suggested Reading and Additional Resources |
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149 | (1) |
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150 | (2) |
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152 | (1) |
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152 | (1) |
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9 Detectors, Sampling, and Image Processing and Analysis |
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153 | (28) |
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153 | (6) |
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9.1.1 Photomultipliers (PMTs) |
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153 | (2) |
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155 | (1) |
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156 | (1) |
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9.1.3.1 Charge-Coupled Device (CCD) Cameras |
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156 | (1) |
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9.1.3.2 Electron-Multiplying CCD (EMCCD) Cameras |
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157 | (1) |
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9.1.3.3 Scientific (sCMOS) Cameras |
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158 | (1) |
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9.2 Noise and the Image Signal-to-Noise Ratio (SNR) |
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159 | (2) |
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161 | (2) |
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9.4 Introduction to Digital Image Processing and Analysis |
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163 | (18) |
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163 | (1) |
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9.4.1.1 Image Restoration |
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164 | (7) |
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9.4.1.2 Image Enhancement |
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171 | (2) |
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9.4.1.3 Binary Conversion/Image Segmentation |
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173 | (1) |
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173 | (3) |
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Suggested Reading and Additional Resources |
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176 | (1) |
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176 | (2) |
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178 | (1) |
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A9.1 Noise and Resolution |
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178 | (1) |
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179 | (1) |
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180 | (1) |
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Section II Medical Imaging |
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181 | (120) |
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183 | (28) |
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10.1 Essence of the Technique |
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183 | (1) |
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183 | (1) |
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10.3 Generating and Detecting US |
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184 | (2) |
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10.4 Transducer Design and Beam Attributes |
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186 | (1) |
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187 | (6) |
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10.5.1 Display of Ultrasound Data |
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191 | (1) |
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191 | (1) |
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191 | (1) |
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191 | (1) |
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192 | (1) |
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10.5.2.1 Two-Dimensional Imaging |
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192 | (1) |
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10.5.2.2 Three-Dimensional Imaging |
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193 | (1) |
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193 | (3) |
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194 | (1) |
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194 | (1) |
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195 | (1) |
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196 | (3) |
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10.7.1 Specular Reflection |
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196 | (1) |
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10.7.2 Nonspecular (Diffuse) Reflection and Scattering |
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196 | (1) |
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197 | (1) |
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10.7.4 Contrast-Enhanced Ultrasound (CEUS) |
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197 | (2) |
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199 | (4) |
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203 | (1) |
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204 | (7) |
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Suggested Reading and Additional Resources |
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206 | (1) |
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207 | (2) |
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209 | (1) |
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A10.1 Diagnostic Ultrasound Imaging of the Carotid Artery |
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209 | (2) |
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11 Projection Radiography and Computed Tomography |
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211 | (32) |
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11.1 Essence of Radiography and CT |
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211 | (1) |
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11.2 Properties of X-Rays |
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211 | (6) |
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11.2.1 Energies of Diagnostic X-Rays |
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211 | (3) |
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11.2.2 Interactions with Matter |
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214 | (1) |
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214 | (3) |
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11.2.2.2 The Compton Effect |
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217 | (1) |
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11.3 Generating and Detecting X-Rays |
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217 | (4) |
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217 | (2) |
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219 | (2) |
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11.4 Projection Radiographic Imaging |
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221 | (6) |
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221 | (1) |
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11.4.1.1 Monoenergetic Beams |
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221 | (2) |
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11.4.1.2 Polyenergetic Beams and Beam Hardening |
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223 | (1) |
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11.4.2 Resolution and Magnification |
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224 | (1) |
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225 | (2) |
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227 | (1) |
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227 | (9) |
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228 | (2) |
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230 | (1) |
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231 | (1) |
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11.5.4 Image Reconstruction |
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231 | (5) |
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236 | (1) |
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236 | (7) |
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Suggested Reading and Additional Resources |
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238 | (1) |
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238 | (2) |
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240 | (1) |
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A11.1 Transmission Imaging (Projection and Tomographic) |
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240 | (3) |
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12 Planar Scintigraphy and Emission Tomography |
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243 | (28) |
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12.1 Essence of Emission Imaging |
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243 | (1) |
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12.2 Physics of Radioactive Materials |
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243 | (5) |
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12.2.1 Atomic and Nuclear Structure |
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243 | (1) |
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12.2.2 Stable and Unstable Nuclei |
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244 | (1) |
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12.2.2.1 Isomeric Transitions |
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244 | (2) |
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12.2.2.2 Positron Emission |
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246 | (1) |
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247 | (1) |
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12.3 Generation and Detection of Radioactivity |
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248 | (4) |
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12.3.1 Generating Radioactivity |
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248 | (1) |
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12.3.2 Detecting Radioactivity |
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249 | (1) |
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12.3.3 Coordinate Computation |
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250 | (1) |
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12.3.4 Pulse Height Analysis |
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251 | (1) |
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12.4 Projection Nuclear Imaging |
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252 | (5) |
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12.4.1 Modern Camera Systems |
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253 | (1) |
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12.4.2 Production of a Digital Image |
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253 | (1) |
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253 | (1) |
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254 | (1) |
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255 | (1) |
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12.4.4 Image Quantification |
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255 | (2) |
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12.5 Emission Computed Tomography |
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257 | (7) |
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258 | (1) |
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12.5.1.1 Data Acquisition |
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258 | (1) |
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258 | (1) |
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259 | (1) |
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12.5.2.1 Data Acquisition |
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259 | (3) |
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262 | (1) |
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12.5.3 Comparison of SPECT and PET |
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262 | (2) |
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264 | (1) |
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264 | (7) |
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Suggested Reading and Additional Resources |
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267 | (1) |
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267 | (2) |
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269 | (1) |
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269 | (2) |
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13 Magnetic Resonance Imaging |
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271 | (30) |
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13.1 Essence of the Technique |
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271 | (1) |
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13.2 Essential Basics from Classical Magnetism and Quantum Mechanics |
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272 | (2) |
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13.2.1 Magnetic Dipole Moments and Angular Momentum |
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272 | (1) |
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13.2.2 Quantum Spin Angular Momentum and Magnetic Moments |
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273 | (1) |
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13.2.3 Energy of a Magnetic Dipole in a Magnetic Field |
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274 | (1) |
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13.2.4 Torque on a Magnetic Dipole in a Magnetic Field |
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274 | (1) |
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13.3 Nuclear Moments and the Strength of the MRI Signal |
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274 | (1) |
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13.4 Overview of an MRI Experiment |
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275 | (1) |
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276 | (2) |
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13.5.1 Quantifying Equilibrium Magnetization |
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276 | (1) |
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13.5.2 Quantum Analysis of RF Fields |
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276 | (2) |
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13.6 An Introduction to Image Production and Quality |
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278 | (5) |
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279 | (1) |
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13.6.1.1 One-Dimensional Imaging |
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280 | (1) |
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13.6.1.2 Two-Dimensional Imaging |
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280 | (1) |
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13.6.2 Image Reconstruction via Fourier Transformation |
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281 | (1) |
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282 | (1) |
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13.7 A More Advanced Analysis |
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283 | (8) |
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13.7.1 Relaxation Mechanisms |
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284 | (1) |
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13.7.1.1 Qualitative Analysis of Relaxation Effects |
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284 | (1) |
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13.7.1.2 Quantifying Transverse Relaxation |
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285 | (1) |
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13.7.1.3 Quantifying Longitudinal Relaxation |
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286 | (1) |
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13.7.2 An Introduction to the Spin Echo |
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287 | (1) |
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288 | (3) |
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13.8 Summary of Two-Dimensional Spin-Echo Imaging |
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291 | (2) |
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13.9 K-Space and the MRI Imaging Equation [ Optional] |
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293 | (2) |
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295 | (6) |
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Suggested Reading and Additional Resources |
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296 | (1) |
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296 | (3) |
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299 | (1) |
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A13.1 Simulating MRI Fourier Data |
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299 | (1) |
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A13.2 Spatial Filtering of MRI Data |
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299 | (1) |
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300 | (1) |
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301 | (4) |
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Appendix A List of Abbreviations |
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301 | (1) |
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Appendix B Fundamental Constants |
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302 | (1) |
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302 | (1) |
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303 | (1) |
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Appendix E Mathematical Relations |
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303 | (2) |
Index |
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305 | |