Fluorescence Spectroscopy and Microscopy, Softcover reprint of the original 1st ed. 2014
Methods and Protocols
Methods in Molecular Biology Series, Vol. 1076

Reflecting the expanding field?s need for reliable protocols, Fluorescence Spectroscopy and Microscopy: Methods and Protocols offers techniques from a worldwide team of experts on this versatile and vital subject.  The topics covered fall into four broad categories: steady-state fluorescence spectroscopy, time-resolved fluorescence spectroscopy, fluorescent probe development, and the various sub-categories of fluorescence microscopy, such as fluorescence recovery after photobleaching (FRAP), live cell FRET imaging (FRETim), fluorescence lifetime imaging (FLIM), fluorescence fluctuation spectroscopy (FFS), and single-molecule fluorescence spectroscopy (smFS).  Written as a part of the popular Methods in Molecular Biology series, chapters include the kind of unambiguous detail and key implementation advice that proves essential for successful results.

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 Fluorescence Imaging Spectroscopy

            Martijn H.W. Stopel, Christian Blum, and Vinod Subramaniam

Part VII: Fluorescence Microscopy: Fluorescence Fluctuation Spectroscopy (FFS)

24. Application of Fluorescence Correlation Spectroscopy (FCS) to Measure the Dynamics of Fluorescent Proteins in Living Cells

            Thomas Weidemann

25. Fluorescence Cross-Correlation Spectroscopy (FCCS) in Living Cells

            Xiaoxiao Ma, Yong Hwee Foo, and Thorsten Wohland

26. Quantifying Lipid-Protein Interaction by Fluorescence Correlation Spectroscopy (FCS)

            Ana M. Melo, Manuel Prieto, and Ana Coutinho

27. PET-FCS: Probing Rapid Structural Fluctuations of Proteins and Nucleic Acids by Single-Molecule Fluorescence Quenching

            Markus Sauer and Hannes Neuweiler

28. Z-Scan Fluorescence Correlation Spectroscopy as a Tool for Diffusion Measurements in Planar Lipid Membranes

            Tomáš Steinberger, Radek Macháň, and Martin Hof

29. Scanning Fluorescence Correlation Spectroscopy (SFCS) with a Scan Path Perpendicular to the Membrane Plane

            Paul Müller, Petra Schwille, and Thomas Weidemann

30. Implementation and Application of Pulsed Interleaved Excitation for Dual-Color FCS and RICS

            Jelle Hendrix and Don C. Lamb

31. Quantitative Study of Protein-Protein Interactions in Live Cell by Dual Color Fluorescence Lifetime Cross-Correlation Spectroscopy

            Sergi Padilla-Parra, Nicolas Audugé, Maïté Coppey-Moisan, and Marc Tramier

32. Brightness Experiments

            Patrick J. Macdonald, Jolene Johnson, Yan Chen, and Joachim D. Mueller

33. Global Analysis of Autocorrelation Functions and Photon Counting Distributions in Fluorescence Fluctuation Spectroscopy

            Victor V. Skakun, Anatoli V. Digris, and Vladimir V. Apanasovich

34. Simulation of Autocorrelation Function and Photon Counting Distribution in Fluorescence Fluctuation Spectroscopy

            Igor P. Shingaryov, Victor V. Skakun, and Vladimir V. Apanasovich

Part VIII: Fluorescence Microscopy: Single-Molecule Fluorescence Spectroscopy (smFS)

35. Single-Molecule Fluorescence of Nucleic Acids

            Kaley McCluskey, Euan Shaw, Daniel A. Lafontaine, and J. Carlos Penedo

36. Photoswitchable Fluorescent Proteins for Superresolution Fluorescence Microscopy Above the Diffraction Limit of Light

            Susana Rocha, Herlinde De Keersmaecker, Hiroshi Uji-i, Johan Hofkens, and Hideaki Mizuno

Explores the methods and applications in the expanding fluorescence field

Provides step-by-step detail essential for reproducible results

Contains key notes and implementation advice from the experts