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Intro
Preface
Contents
1: The Physical Principles of Light Propagation and Light-Matter Interactions
1.1 Definitions of Light
1.2 Particle and Wave Properties of Light
1.3 Polarization
1.4 Refraction
1.5 Reflection
1.6 Light Scattering
1.7 Diffraction and Its Limits
1.8 Photon Absorption, Fluorescence, and Stimulated Emission
1.9 Filtering Light
References
Further Reading
2: Design, Alignment, and Usage of Infinity-Corrected Microscope
2.1 Development of Compound Microscope with Infinity Optics

2.2 Design Parameters: Resolution Limit, Numerical Aperture, Magnification, Depth of Field, and Field of View
2.3 Optical Aberrations and Their Corrections
2.4 Design Specifications of the Infinity-Corrected Microscope Objective
2.5 Critical and Köhler Illuminations
2.6 Components of Infinity-Corrected Microscope System
2.7 Alignment for Designing the Infinity-Corrected Bright Field Microscopy
2.8 Label-Free and Quantitative Phase Microscopy
2.8.1 Dark Field Microscopy
2.8.2 Zernike Phase Contrast Microscopy
2.8.3 Differential Interference Contrast (DIC) Microscope

2.8.4 Digital Holographic Microscopy for Quantitative Phase Measurement
References
3: Epifluorescence Microscopy
3.1 Fluorescence
3.1.1 Jablonski Diagram
3.1.2 Fluorescent Markers
3.2 Fluorescence Microscope Setup
3.2.1 Excitation Module
3.2.2 Objective
3.2.3 Fluorescence Filters
3.2.4 Emission Module
3.3 Basics of Sample Preparation for Fluorescence Microscopy
3.4 Main Applications of Fluorescence Microscopy
3.4.1 Structural Imaging
3.4.2 Functional Imaging
References
Further Reading

4: Basic Digital Image Acquisition, Design, Processing, Analysis, Management, and Presentation
4.1 Image Acquisition and Analysis: Why they Are Needed
4.1.1 Image Relevance in Science from Biology to Astronomy
4.1.2 Need for Quantitative Methods Is Extended to Light Microscopy
4.2 Representation of Reality: Image Acquisition
4.2.1 What Is the Difference Between a Material Object and a Representation of It?
4.2.2 Light Microscopy Key Concepts
4.2.2.1 What Is Light Microscopy?
4.2.2.2 Optical Resolution
4.2.2.3 Microscope Elements
4.2.2.4 Fluorescence

4.2.2.5 Transmitted Light
4.2.2.6 Detectors: Cameras and PMT/Hybrid
4.2.2.7 Digitization
4.2.3 Examples of Microscope Systems: Simplified Light Path for Wide-Field and Confocal Laser Scanning Microscopy
4.2.4 Experiment Design
4.2.5 Questions and Answers
4.3 Image Visualization Methods
4.4 Image Analysis
4.4.1 Questions and Answers
4.4.2 Questions and Answers
4.5 Publication of Images and Data
4.5.1 Questions and Answers
References
Additional References
5: Confocal Microscopy
5.1 Introduction
5.2 Fluorescence Blurring

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