Quantitative Fluorescence Microscopy Techniques

  • Alessandro Esposito
  • Simon Schlachter
  • Gabriele S. Kaminski Schierle
  • Alan D. Elder
  • Alberto Diaspro
  • Fred S. Wouters
  • Clemens F. Kaminski
  • Asparouh I. Iliev
Part of the Methods in Molecular Biology book series (MIMB, volume 586)


Fluorescence microscopy is a non-invasive technique that allows high resolution imaging of cytoskeletal structures. Advances in the field of fluorescent labelling (e.g., fluorescent proteins, quantum dots, tetracystein domains) and optics (e.g., super-resolution techniques and quantitative methods) not only provide better images of the cytoskeleton, but also offer an opportunity to quantify the complex of molecular events that populate this highly organised, yet dynamic, structure.

For instance, fluorescence lifetime imaging microscopy and Förster resonance energy transfer imaging allow mapping of protein–protein interactions; furthermore, techniques based on the measurement of photobleaching kinetics (e.g., fluorescence recovery after photobleaching, fluorescence loss in photobleaching, and fluorescence localisation after photobleaching) permit the characterisation of axonal transport and, more generally, diffusion of relevant biomolecules.

Quantitative fluorescence microscopy techniques offer powerful tools for understanding the physiological and pathological roles of molecular machineries in the living cell.

Key words




AE is funded by the Engineering and Physical Sciences Research Council (EPSRC, EP/F044011/1), UK. AII is financed by the Emmy Noether Programme of the German Science Foundation (Deutsche Forschungsgemeinschaft (DFG)) and the DFG-funded Rudolf Virchow Zentrum for Experimental Medicine at the University of Würzburg, Germany. CFK is funded by the Leverhulme Trust (UK), the Biotechnology and Biological Sciences Research Council (BBSRC, UK) and the EPSRC. FSW is financed by the Excellence Cluster 171 “Microscopy on the Nanometer Scale” of the DFG-funded Center for Molecular Physiology of the Brain (CMPB) and by the German Federal Ministry for Education and Research (BMBF). AD is funded by the University of Genoa and the Ministry of Education, University and Research (MIUR).


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Copyright information

© Humana Press, a part of Springer Science+Business Media, LLC 2009

Authors and Affiliations

  • Alessandro Esposito
    • 1
  • Simon Schlachter
    • 1
  • Gabriele S. Kaminski Schierle
    • 1
  • Alan D. Elder
    • 1
  • Alberto Diaspro
    • 2
  • Fred S. Wouters
    • 3
  • Clemens F. Kaminski
    • 1
  • Asparouh I. Iliev
    • 4
  1. 1.Department of Chemical Engineering and BiotechnologyUniversity of CambridgeCambridgeUK
  2. 2.LAMBS-MicroScoBio, Department of PhysicsUniversity of GenoaGenoaItaly
  3. 3.Laboratory for Molecular and Cellular Systems, Department of Neuro- and Sensory PhysiologyUniversity Medicine GöttingenGöttingenGermany
  4. 4.Rudolf Virchow Center & Institute of Pharmacology and ToxicologyUniversity of WürzburgWürzburgGermany

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