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
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 586)

Summary

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

FRET FLIM FRAP FLAP FLIP Bioimaging 

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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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