Part I: Steady-State Fluorescence Spectroscopy (SSFS)

1. How to Collect National Institute of Standards and Technology (NIST) Traceable Fluorescence Excitation and Emission Spectra

Adam Matthew Gilmore

2. Steady State Fluorescence Polarization/Anisotropy for the Study of Protein Interactions

Nicholas G. James and David M. Jameson

3. Quantitative Fluorescence Spectral Analysis of Protein Denaturation

Ivo H.M. van Stokkum and Sergey P. Laptenok

4. High-Pressure Fluorescence Applications

Mariano Dellarole and Catherine A. Royer

Part II: Time-Resolved Fluorescence Spectroscopy (TRFS)

5. Frequency Domain Fluorometry: Theory and Application

Carissa M. Vetromile and David M. Jameson

6. Polar Plot Representation of Time-Resolved Fluorescence

John Paul Eichorts, Kai wen Teng, and Robert M. Clegg

7. Ensemble and Single Molecule Detected Time Resolved FRET Methods in Studies of Protein Conformations and Dynamics

Tomer Orevi, Eitan Lerner, Gil Rahamim, Dan Amir, and Elisha Haas

8. MD + QM Correlations with Tryptophan Fluorescence Spectral Shifts and Lifetimes

Patrik R. Callis and Jose R. Tusell

9. Analysis of Time-Dependent Red Shifts in Fluorescence Emission from Tryptophan Residues in Proteins

Dmitri Toptygin

10. Global Analysis of Time-Resolved Fluorescence Data

Anatoli V. Digris, Eugene G. Novikov, Victor V. Skakun, and Vladimir V. Apanasovich

11. Nanometrology

David J.S. Birch and Philip Yip

12. Upconversion Spectrophotofluorometry

Arianna Biesso, Jianhua Xu, and Jay R. Knutson

13. Subpicosecond Kerr-Gate Spectrofluorometry

Sergey P. Laptenok, Patrick Nürnberger, Andras Lukacs, and Marten H. Vos

14. Photo-Induced Electron Transfer Modeling to Simulate Flavoprotein Fluorescence Decay

Nadtanet Nunthaboot, Kiattisak Lugsanangarm, Arthit Nueangaudom, Somsak Pianwanit, Sirirat Kokpol, and Fumio Tanaka

Part III: Fluorescent Probe Development (FPD)

15. Biosynthetic Incorporation of Trp Analogs in Proteins

Jaap Broos

16. Optimization of Fluorescent Proteins

Daphne S. Bindels, Joachim Goedhart, Mark A. Hink, Laura van Weeren, Linda Joosen, and Theodorus W.J. Gadella, Jr.

17. Monitoring Membrane Properties and Apoptosis Using Membrane Probes of the 3-Hydroxyflavone Family

Zeinab Darwich, Andrey S. Klymchenko, and Yves Mély

Part IV: Fluorescence Microscopy: Fluorescence Recovery After Photobleaching (FRAP)

18. Rectangle FRAP for Measuring Diffusion with a Laser Scanning Microscope

Ranhua Xiong, Hendrik Deschout, Jo Demeester, Stefaan C. De Smedt, and Kevin Braeckmans

Part V: Fluorescence Microscopy: Förster Resonance Energy Transfer Imaging (FRETim)

19. A Quantitative Protocol for Intensity-Based Live Cell FRET Imaging

Clemens F. Kaminski, Eric. J. Rees, and Gabriele S. Kaminski Schierle

Part VI: Fluorescence Microscopy: Fluorescence Lifetime Imaging (FLIM)

20. Widefield Fluorescence Lifetime Imaging with Multi-Anode Detectors

Roland Hartig, Yury Prokazov, Evgeny Turbin, and Werner Zuschratter

21. Global Analysis of FRET- FLIM Data in Live Plant Cells

Sergey P. Laptenok, Joris J. Snellenburg, Christoph A. Bücherl, Kai R. Konrad, and Jan Willem Borst

22. Time-Resolved Fluorescence Anisotropy Imaging

Klaus Suhling, James Levitt, and Pei-Hua Chung

23. Multi-Modal Fluo