Fluorescence cross-correlation spectroscopy in living cells

Nat Methods. 2006 Feb;3(2):83-9. doi: 10.1038/nmeth822.


Cell biologists strive to characterize molecular interactions directly in the intracellular environment. The intrinsic resolution of optical microscopy, however, allows visualization of only coarse subcellular localization. By extracting information from molecular dynamics, fluorescence cross-correlation spectroscopy (FCCS) grants access to processes on a molecular scale, such as diffusion, binding, enzymatic reactions and codiffusion, and has become a valuable tool for studies in living cells. Here we review basic principles of FCCS and focus on seminal applications, including examples of intracellular signaling and trafficking. We consider FCCS in the context of fluorescence resonance energy transfer and multicolor imaging techniques and discuss application strategies and recent technical advances.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Algorithms
  • Animals
  • Biological Transport
  • Diffusion
  • Endocytosis / physiology
  • Enzymes / metabolism
  • Fluorescence Resonance Energy Transfer / methods
  • Humans
  • Laser Scanning Cytometry / instrumentation
  • Laser Scanning Cytometry / methods*
  • Oligonucleotides / metabolism
  • Protein Binding
  • Protein Transport
  • Proteins / metabolism
  • Signal Transduction / physiology
  • Spectrometry, Fluorescence / instrumentation
  • Spectrometry, Fluorescence / methods


  • Enzymes
  • Oligonucleotides
  • Proteins