Digital Microscopy

The previous edition of this book marked the shift in technology from video to digital camera use with microscope use in biological science. This new edition presents some of the optical fundamentals needed to provide a quality image to the digital camera. Specifically, it covers the fundamental geometric optics of finite- and infinity-corrected microscopes, develops the concepts of physical optics and Abbe’s theory of image formation, presents the principles of Kohler illumination, and finally reviews the fundamentals of fluorescence and fluorescence microscopy. The second group of chapters deals with digital and video fundamentals: how digital and video cameras work, how to coordinate cameras with microscopes, how to deal with digital data, the fundamentals of image processing, and low light level cameras. The third group of chapters address some specialized areas of microscopy that allow sophisticated measurements of events in living cells that are below the optical limits of resolution.

Expands coverage to include discussion of confocal microscopy not found in the previous editionIncludes "traps and pitfalls" as well as laboratory exercises to help illustrate methods

Microscope Basics.
The Optics of Microscope Image Formation.
Proper Alignment of the Microscope.
Mating Cameras to Microscopes.
Fundamentals of Fluorescence and Fluorescence Microscopy.
Fluorescent Protein Applications in Microscopy.
Live Cell Fluorescence Imaging.
Working with Classic Video.
Practical Aspects of Adjusting Digital Cameras.
Cameras for Digital Microscopy.
A High-Resolution Multimode Digital Microscope System.
Electronic Cameras for Low-Light Microscopy.
Camera Technologies for Low Light Imaging: Overview and Relative Advantages.
Digital Manipulation of Brightfield and Fluorescence Images: Noise Reduction, Contrast Enhancement, and Feature Extraction.
Digital Image Files in Light Microscopy.
High-Resolution Video–Enhanced Differential Interference Contrast Light Microscopy.
Quantitative Analysis of Digital Microscope Images.
Evaluating Optical Aberration Using Fluorescent Microspheres: Methods, Analysis, and Corrective Actions.
Ratio Imaging: Practical Considerations for Measuring Intracellular Ca++ and pH in Living Cells.
Computational Restoration of Fluorescence Images: Noise Reduction, Deconvolution, and Pattern Recognition.
Quantitative Fluorescence Microscopy and Image Deconvolution.
Practical Aspects of Quantitative Confocal Microscopy.
Theoretical Principles and Practical Considerations for Fluorescence Resonance Energy Transfer Microscopy.
Fluorescence Lifetime Imaging Microscopy.
Fluorescence Correlation Spectroscopy: Molecular Complexing in Solution and in Living Cells.
Breaking the Resolution Limit in Light Microscopy.