Optical Imaging Techniques in Cell Biology

This reference on optical imaging techniques covers the field from the basics of the optical microscope to highly advanced techniques such as Förster resonant energy transfer (FRET) and fluorescence lifetime imaging microscopy (FLIM). Using a uniform style and format, it provides an understanding of the underlying principles, so readers can learn the benefits and limitations of each technique described. Topics covered include: optical contrasting techniques, fluorescence and fluorescence microscopes, three-dimensional imaging, the confocal microscope, nonlinear microscopy, advanced fluorescence techniques, and breaking the diffraction limit. Chapters include suggestions for further reading.

The Light Microscope. Optical Contrasting Techniques. Fluorescence and Fluorescence Microscopy. Image Capture. The Confocal Microscope. The Digital Image. Aberrations and Their Consequences. Nonlinear Microscopy. High-Speed Confocal Microscopy. Deconvolution and Image Processing. Three-Dimensional Imaging: Stereoscopy and Reconstruction. Green Fluorescent Protein. Fluorescent Staining. Quantitative Fluorescence. Advanced Fluorescence Techniques: FLIM, FRET, and FCS. Evanescent Wave Microscopy. Beyond the Diffraction Limit. Appendix A: Microscope Care and Maintenance. Appendix B: Keeping Cells Alive under the Microscope. Appendix C: Antibody Labeling of Plant and Animal Cells: Tips and Sample Schedules. Appendix D: Image Processing with ImageJ.