Fluorescence Spectroscopy and Microscopy

Methods and Protocols

  • Yves Engelborghs
  • Antonie J.W.G. Visser

Part of the Methods in Molecular Biology book series (MIMB, volume 1076)

Table of contents

  1. Front Matter
    Pages i-xxi
  2. Steady-State Fluorescence Spectroscopy (SSFS)

  3. Time-Resolved Fluorescence Spectroscopy (TRFS)

    1. Front Matter
      Pages 75-75
    2. Carissa M. Vetromile, David M. Jameson
      Pages 77-95
    3. John Paul Eichorst, Kai Wen Teng, Robert M. Clegg
      Pages 97-112
    4. Tomer Orevi, Eitan Lerner, Gil Rahamim, Dan Amir, Elisha Haas
      Pages 113-169
    5. Anatoli V. Digris, Eugene G. Novikov, Victor V. Skakun, Vladimir V. Apanasovich
      Pages 257-277
    6. David J. S. Birch, Philip Yip
      Pages 279-302
    7. Arianna Biesso, Jianhua Xu, Jay R. Knutson
      Pages 303-319
    8. Sergey P. Laptenok, Patrick Nuernberger, Andras Lukacs, Marten H. Vos
      Pages 321-336
    9. Nadtanet Nunthaboot, Kiattisak Lugsanangarm, Arthit Nueangaudom, Somsak Pianwanit, Sirirat Kokpol, Fumio Tanaka
      Pages 337-355
  4. Fluorescent Probe Development (FPD)

    1. Front Matter
      Pages 357-357
    2. Daphne S. Bindels, Joachim Goedhart, Mark A. Hink, Laura van Weeren, Linda Joosen, Theodorus W. J. Gadella Jr.
      Pages 371-417
    3. Zeinab Darwich, Andrey S. Klymchenko, Yves Mély
      Pages 419-430
  5. Fluorescence Microscopy: Fluorescence Recovery After Photobleaching (FRAP)

    1. Front Matter
      Pages 431-431
    2. Ranhua Xiong, Hendrik Deschout, Jo Demeester, Stefaan C. De Smedt, Kevin Braeckmans
      Pages 433-441
  6. Fluorescence Microscopy: Förster Resonance Energy Transfer Imaging (FRETim)

    1. Front Matter
      Pages 443-443
    2. Clemens F. Kaminski, Eric J. Rees, Gabriele S. Kaminski Schierle
      Pages 445-454
  7. Fluorescence Microscopy: Fluorescence Lifetime Imaging (FLIM)

    1. Front Matter
      Pages 455-455
    2. Roland Hartig, Yury Prokazov, Evgeny Turbin, Werner Zuschratter
      Pages 457-480
    3. Sergey P. Laptenok, Joris J. Snellenburg, Christoph A. Bücherl, Kai R. Konrad, Jan Willem Borst
      Pages 481-502
    4. Klaus Suhling, James Levitt, Pei-Hua Chung
      Pages 503-519
    5. Martijn H. W. Stopel, Christian Blum, Vinod Subramaniam
      Pages 521-536
  8. Fluorescence Microscopy: Fluorescence Fluctuation Spectroscopy (FFS)

    1. Front Matter
      Pages 537-537
    2. Xiaoxiao Ma, Yong Hwee Foo, Thorsten Wohland
      Pages 557-573
    3. Ana M. Melo, Manuel Prieto, Ana Coutinho
      Pages 575-595
    4. Sergi Padilla-Parra, Nicolas Audugé, Maïté Coppey-Moisan, Marc Tramier
      Pages 683-698
    5. Patrick J. Macdonald, Jolene Johnson, Yan Chen, Joachim D. Mueller
      Pages 699-718
    6. Igor P. Shingaryov, Victor V. Skakun, Vladimir V. Apanasovich
      Pages 743-755
  9. Fluorescence Microscopy: Single-Molecule Fluorescence Spectroscopy (smFS)

    1. Front Matter
      Pages 757-757
    2. Kaley McCluskey, Euan Shaw, Daniel A. Lafontaine, J. Carlos Penedo
      Pages 759-791
    3. Susana Rocha, Herlinde De Keersmaecker, Hiroshi Uji-i, Johan Hofkens, Hideaki Mizuno
      Pages 793-812
  10. Back Matter
    Pages 813-816

About this book


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.


Comprehensive and practical, Fluorescence Spectroscopy and Microscopy: Methods and Protocols aims to guide both ‘novice’ and established scientists toward furthering their research with these invaluable techniques.


Cuvette-based studies Fluorescence Fluctuation Spectroscopy Fluorescence Lifetime Imaging Fluorescence Recovery After Photobleaching Fluorescent Probe Development Förster Resonance Energy Transfer Imaging Green fluorescent protein (GFP) Single-Molecule Fluorescence Spectroscopy Steady-State Fluorescence Spectroscopy Time-Resolved Fluorescence Spectroscopy

Editors and affiliations

  • Yves Engelborghs
    • 1
  • Antonie J.W.G. Visser
    • 2
  1. 1.KULeuven-Biomolecular Dynamics BioSCENTerLeuvenBelgium
  2. 2.MicroSpectroscopy CentreUniversity of Wageningen BiochemistryWageningenThe Netherlands

Bibliographic information

  • DOI
  • Copyright Information Springer Science+Business Media, LLC 2014
  • Publisher Name Humana Press, Totowa, NJ
  • eBook Packages Springer Protocols
  • Print ISBN 978-1-62703-648-1
  • Online ISBN 978-1-62703-649-8
  • Series Print ISSN 1064-3745
  • Series Online ISSN 1940-6029
  • Buy this book on publisher's site