This book offers a complete presentation of the physics, math, and experimental setups for both TIRF and related evanescence microscopies. It covers evanescence in both fluorescence excitation or emission. It also discusses, in detail, the theory, setups, and practical biological/biochemical applications for combinations of evanescence microscopies with other optical techniques such as polarization, photobleaching, correlation spectroscopy, scattering, image enhancement, optical force, two-photon, energy transfer, structured illumination, scanning, quenching, and single molecule detection. Physical and qualitative discussions augment the rigorous math, making the book accessible and interesting to a wide range of audiences with backgrounds from biology to chemistry to physics. The book also contains useful step-by-step guides to building, modifying, and aligning TIRF microscopy systems for specialized purposes.

This book is suitable for researchers who work with TIRF and near field optics as a standard technique, as well as those who want to develop their techniques further. Students in graduate level optics, microscopy course, and advanced biophysics courses will also find this book useful.

Daniel Axelrod is a Professor Emeritus of Physics and Biophysics at the University of Michigan, Ann Arbor. His specialty is developing novel optical microscopy techniques to study the motion and organization of biological molecules and cellular organelles near biological surfaces. Many biological aspects are done in collaboration with research groups at the University of Michigan Medical School. Professor Axelrod is a Fellow of the Biophysical Society and a recipient of the Biophysical Society's 2010 Gregorio Weber Award for fluorescence theory and applications.