Fluorescence correlation spectroscopy (FCS)

FCS is a method to detect molecular parameters like e.g. diffusion time, diffusion coefficent. Threrefore FCS can be used to analyse interaction of molecules (Elson & Magde, 1974; Magde et al., 1974; overview: Schwille & Haustein, 2001). In Fluorescence Correlation Spectroscopy the fluctuation of the fluorescence intensity is recorded in a very small volume, i.e. in femtoliter range via a confocal optic system.

Fluorescence Intensity Distribution Analysis

The photon statistic method FIDA (Fluorescence Intensity Distribution Analysis) was developed in 1999 by Kask (Kask, P., Palo, K., Ullman, D., and Gall, K. (1999) Proc. Natl. Acad. Sci. U. S. A. 96, 13756-13761) and in parallel by Chen et al. (1999) Biophys. J. 77, 553-567, where it was named PCH (Photon Count Histogram).

The method FIDA was quickly developed into a family of photon statistic methods (2D-FIDA, Kask, P., Palo, K., Fay, N., Brand, L., Mets, Ü., Ullman, D., Jungmann, J., Pschorr, J., and Gall, K. (2000) Biophys. J. 78, 1703-1713, FIMDA, , Palo K., Mets, Ü., Jäger, S., Kask, P., and Gall, K. (2000) Biophys. J. 79, FILDA, Palo, K., Brand, L., Eggeling, C., Jäger, S., Kask, P., and Gall, K. (2002) Biophys. J. 83, 605-618). In all three methods additional information (2nd detector in 2D-FIDA, molecular diffusion in FIMDA and fluorescence lifetime in FILDA) is used to improve statistical accuracy and/or gain additional information. All these methods deliver detailed information about a fluorescent tracer diffusing through the open 3D volume of the size of one bacteria (roughly one femto-liter). As long as enough fluorescent molecules are available (nano-molar range) this methods are well suited for diagnostic or drug discovery.

  • Goossens K, Prior M, Pacheco V, Willbold D, Müllen K, Enderlein J, Hofkens J, Gregor I 
    Accurate diffusion coefficients of organosoluble reference dyes in organic media measured by dual-focus fluorescence correlation spectroscopy. 
    ACS Nano 9, 7360-7373 (2015) 
  • Bannach O, Reinartz E, Henke F, Dreßen F, Oelschlegel A, Kaatz M, Groschup M H, Willbold D, Riesner D and Birkmann E 
    Analysis of prion protein aggregates in blood and brain from pre-clinical and clinical BSE cases 
    Vet Microbiol 166, 102-108 (2013) 
  • Hickman DT, López-Deber MP, Ndao DM, Silva AB, Nand D, Pihlgren M, Giriens V, Madani R, St-Pierre A, Karastan H, Nagel-Steger L, Willbold D, Riesner D, Nicolau C, Baldus M, Pfeifer A, Muhs A 
    Sequence-independent control of peptide conformation in liposomal vaccines for targeting protein misfolding diseases. 
    J. Biol.Chem. 286, 13966-13976 (2011) 
  • Nagel-Steger L, Demeler B, Meyer-Zaika W, Hochdörffer K, Schrader T, Willbold D 
    Modulation of aggregate size- and shape-distributions of the amyloid-beta peptide by a designed ß-sheet breaker. 
    Eur. Biophys. J. 39, 415-422 (2010) 
  • Funke SA, Birkmann E, Willbold D 
    Detection of Amyloid-β aggregates in body fluids: A suitable method for early diagnosis of Alzheimer's diesease? 
    Curr. Alzheimer Res. 6, 285-289 (2009) 
  • Birkmann E, Henke F, Funke SA, Bannach O, Riesner D, Willbold D 
    A highly sensitive diagnsotic assay for aggregate-related diseases e.g. prion disease and Alzheimer's disease. 
    Rejuvenation Res. 11, 359-363 (2008) 
  • Funke SA, Birkmann E, Henke F, Görtz P, Lange-Asschenfeldt C, Riesner D, Willbold D 
    An ultra sensitive assay for early diagnosis of Alzheimer's disease. 
    Rejuvenation Res. 11, 315-318 (2008) 
  • Mueller CB, Loman A, Pacheco V, Koberling F, Willbold D, Richtering W, Enderlein J 
    Precise measurement of diffusion by multi-color dual-focus fluorescence correlation spectroscopy. 
    Europhys. Lett. 83, 46001 (2008) 
  • Nagel-Steger L, Demeler B, Hochdörfer K, Schrader T, Willbold D 
    Modulation of aggregate size and shape distributions of amyloid-ß peptide solutions by a designed ß-sheet breaker. 
    NIC Workshop 2008 (From Computational Biophysics to Systems Biology; CBSB08) Vol. 40, (2008) 
  • Birkmann E, Henke F, Weinmann N, Dumpitak C, Groshup M, Funke SA, Willbold D, Riesner D, 
    Counting of single prion particles bound to a capture-antibody surface (surface-FIDA), 
    Vet. Microbiol. 123, 294-304 (2007) 
  • Elfrink K, Nagel-Steger L, Riesner D 
    Interaction of the cellular prion protein with raft-like lipid membranes. 
    Biol. Chem. 388, 79-90 (2007) 
    http://dx.doi.org/10.1515/BC.2007.010, 01/01/2007  
  • Funke SA, Birkmann E, Henke F, Görtz P, Lange-Asschenfeldt C, Riesner D, Willbold D, 
    Single particle detection of amyloid-ß aggregates associated with Alzheimer's disease. 
    Biochem. Biophys. Res. Comm. 364, 902-907 (2007) 
  • Birkmann E, Schafer O, Weinmann N, Dumpitak C, Beekes M, Jackman R, Thorne L, Riesner D 
    Detection of prion particles in samples of BSE and scrapie by fluorescence correlation spectroscopy without proteinase K digestion. 
    Biol Chem. 387, 95-102 (2006) 
  • Leffers KW, Schell J, Jansen K, Lucassen R, Kaimann T, Nagel-Steger L, Tatzelt J, Riesner D 
    The structural transition of the prion protein into its pathogenic conformation is induced by unmasking hydrophobic sites. 
    J Mol Biol 344, 839-853 (2004) 

Physikalische Biologie
Gebäude 26.12, Ebene U1
Universitätsstraße 1
40225 Düsseldorf

Prof. Dr. D. Willbold
Gebäude 26.12
Ebene U1, Raum 84
Telefon: +49-(0)211 / 81-11390
Mail senden

Verantwortlich für den Inhalt: E-Mail sendenRedaktionsteam IPB