Total internal reflection fluorescence (TIRF) and evanescence microscopies /

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"Version: 20220901"--Title page verso.Includes bibliographical references.1. Introduction to optical evanescence -- 1.1. Overview -- 1.2. Applications to biochemistry and cell biology -- 1.3. Ray picture of total internal reflection -- 1.4. Maxwell's equations and wave numbers -- 1.5. Causes of evanescence : a physical view2. Total internal reflection theory -- 2.1. Rays and TIR -- 2.2. Waves and TIR -- 2.3. Evanescent intensity -- 2.4. Finite-width incident beams : the Goos-H?anchen shift -- 2.5. Reflected intensities3. Structure in the lower-index material -- 3.1. Light absorption in medium 1 -- 3.2. Intermediate layers -- 3.3. Metal films and surface plasmons -- 3.4. Slab waveguides -- 3.5. Total internal reflection scattering4. Emission of fluorophores near a surface -- 4.1. The emission near field : a semi-qualitative view -- 4.2. Capture of the near field : summary of quantitative theory -- 4.3. Polarization of the emitted electric field -- 4.4. Emitted intensity and total power -- 4.5. Emitted intensity vs polar angle -- 4.6. Total fluorescence collection through a microscope objective -- 4.7. Pattern at the back focal plane -- 4.8. Characterization of films with supercritical-emission light -- 4.9. Effect of metal films on fluorescence emission -- 4.10. Pattern at the image plane -- 4.11. Virtual supercritical angle fluorescence microscopy (vSAF) -- 4.12. Emission polarization including supercritical light -- 4.13. SAF/UAF : measurement of the absolute distance between a fluorophore and a surface -- 4.14. Effect of near-field capture on fluorescence lifetime5 Optical configurations and setup -- 5.1. Inverted microscope TIR with prism above -- 5.2. Inverted microscope TIR with prism below -- 5.3. Upright microscope TIR with prism below -- 5.4. Objective-based TIR -- 5.5. Incidence angle, multicolor, and polarization control -- 5.6. Alignment -- 5.7. Rapid chopping between TIR and epi-illumination -- 5.8. Supercritical-angle fluorescence (SAF) emission setup -- 5.9. Imaging the back focal plane directly -- 5.10. Measurement of evanescent field depth -- 5.11. TIRF-structured illumination microscopy (TIRF-SIM)6. Applications of TIRF microscopy and its combination with other fluorescence techniques -- 6.1. Refractive indices in cell cultures -- 6.2. Axial position and motion of cell components -- 6.3. Quenching with a metal film -- 6.4. Image sharpening in TIR -- 6.5. Polarized excitation TIRF -- 6.6. Variable-depth TIRF -- 6.7. Optical force in an evanescent field -- 6.8. TIR/FCS and TIR/FRAP -- 6.9. TIR-continuous photobleaching -- 6.10. TIR-FRET -- 6.11. Two-photon TIRF.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. Part of Biophysical Society-IOP series.Researchers who work with TIRF and near field optics as a standard technique, and those who want to develop the techniques farther.Also available in print.Mode of access: World Wide Web.System requirements: Adobe Acrobat Reader, EPUB reader, or Kindle reader.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.Title from PDF title page (viewed on October 5, 2022).

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Publisher

: .,

Collation

1 online resource (various pagings) :illustrations (some color).

Language

English

ISBN/ISSN

9780750333511

Classification

570.282

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-

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-

Edition

-

Subject(s)

Biophysics.
Medical physics and biophysics.
Fluorescence microscopy.
Total internal reflection (Optics)

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Daniel Axelrod.

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