Abstract

Fluorescence correlation spectroscopy (FCS) is a powerful spectroscopic technique for studying samples at dilute fluorophore concentrations down to single molecules. The standard way of data acquisition, at such low concentrations, is an asynchronous photon counting mode that generates data only when a photon is detected. A significant problem is how to efficiently convert such asynchronously recorded photon count data into a FCS curve. This problem becomes even more challenging for more complex correlation analysis such as the recently introduced combination of FCS and time-correlated single-photon counting (TCSPC). Here, we present, analyze, and apply an algorithm that is highly efficient and can easily be adapted to arbitrarily complex correlation analysis.

© 2003 Optical Society of America

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    [CrossRef]
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    [CrossRef]
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    [CrossRef]
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    [CrossRef]
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    [CrossRef]
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    [CrossRef]
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    [CrossRef]
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    [CrossRef]

Appl. Phys. B. (1)

K. Schätzel �??Correlation techniques in dynamic light scattering,�?? Appl. Phys. B. 42, 193-213 (1987).
[CrossRef]

Biophys. J. (2)

P. Schwille, F.J. Meyer-Almes, R. Rigler �??Dual-color fluorescence cross-correlation spectroscopy for multicomponent diffusional analysis in solution,�?? Biophys. J. 72, 1878-86 (1997).

D.C. Lamb, A. Schenk, C. Röcker, C. Scalfi-Happ, G.U. Nienhaus �??Sensitivity Enhancement in Fluorescence Correlation Spectroscopy of Multiple Species Using Time-Gated Detection,�?? Biophys. J. 79, 1129-38 (2000).

Chem. Phys. Lett. (1)

M. Böhmer, M. Wahl, H.J. Rahn, R. Erdmann, J. Enderlein �??Time-resolved fluorescence correlation spectroscopy,�?? Chem. Phys. Lett. 353, 439-45 (2002).
[CrossRef]

J. Biotechnol. (1)

C. Eggeling, S. Berger, L. Brand, J.R. Fries, J. Schaffer, A. Volkmer, C.A.M. Seidel �??Data registration and selective single-molecule anaylsis using mulit-parameter fluorescence detection,�?? J. Biotechnol. 86, 163-80 (2001).
[CrossRef]

J. Mod. Opt. (1)

K. Schätzel, M. Drewel, S. Stimac �??Photon correlation measurements at large lag times: Improving statistical accuracy,�?? J. Mod. Opt. 35, 711-8 (1988).

Rev. Sci. Instrum. (5)

M. Höbel, J. Ricka �??Dead-time and afterpulsing correction in multiphoton timing with nonideal detectors,�?? Rev. Sci. Instrum. 65, 2326-36 (1994).
[CrossRef]

W. Becker, H. Hickl, C. Zander, K.H. Drexhage, M. Sauer, S. Siebert, J. Wolfrum �??Time-resolved detection and identification of single analyte molecules in microcapillaries by time-correlated single-photon counting (TCSPC),�?? Rev. Sci. Instrum. 70, 1835-41 (1999).
[CrossRef]

M. Böhmer, F. Pampaloni, M. Wahl, H.J. Rahn, R. Erdmann, J. Enderlein �??Advanced Time-Resolved Confocal Scanning Device For Ultrasensitive Fluorescence Detection,�?? Rev. Sci. Instrum. 72, 4145-52 (2001).
[CrossRef]

J.S. Eid, J.D. Müller, E. Gratton �??Data acquisition card for fluctuation correlation spectroscopy allowing full access to the detected photon sequence,�?? Rev. Sci. Instrum. 71, 361-8 (2000).
[CrossRef]

D. Magatti, F. Ferri �??25 ns software correlator for photon and fluorescence correlation spectroscopy,�?? Rev. Sci. Instrum. 74, 1135-44 (2003).
[CrossRef]

Other (3)

M. Böhmer, J. Enderlein �??Single molecule detection on surfaces with the confocal laser scanning microscope,�?? in ref.[1], pp.145-83.

R. Rigler, E. Elson (Eds.) Fluorescence Correlation Spectroscopy (Springer, New York/Berlin, 2001).

C. Zander, J. Enderlein, R.A. Keller (Eds.) Single-Molecule Detection in Solution - Methods and Applications (VCH-Wiley, Berlin/New York, 2002).

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