OSA's Digital Library

Applied Optics

Applied Optics


  • Vol. 42, Iss. 19 — Jul. 1, 2003
  • pp: 4031–4036

Afterpulsing and its correction in fluorescence correlation spectroscopy experiments

Ming Zhao, Lei Jin, Bo Chen, Yao Ding, Hui Ma, and Dieyan Chen  »View Author Affiliations

Applied Optics, Vol. 42, Issue 19, pp. 4031-4036 (2003)

View Full Text Article

Enhanced HTML    Acrobat PDF (117 KB)

Browse Journals / Lookup Meetings

Browse by Journal and Year


Lookup Conference Papers

Close Browse Journals / Lookup Meetings

Article Tools



Afterpulsing arises from feedback in a photon detector. This means that each real signal pulse can be followed by an afterpulse at a later time. This effect is particularly troubling in photon correlation experiments. Few treatments of this effect have appeared in the literature, and few software programs to solve the problem have been written. We demonstrate the afterpulsing effect in fluorescence correlation spectroscopy by using different avalanche photodiodes. We prove theoretically that under simple and reasonable conditions afterpulsing in autocorrelation can be eliminated to the leading order; we have found it easy to program software for the correction. We compare our results with those from cross correlation. We also discuss some experimental parameters that may affect the afterpulsing.

© 2003 Optical Society of America

OCIS Codes
(040.5160) Detectors : Photodetectors
(300.2530) Spectroscopy : Fluorescence, laser-induced

Original Manuscript: July 27, 2002
Revised Manuscript: January 6, 2003
Published: July 1, 2003

Ming Zhao, Lei Jin, Bo Chen, Yao Ding, Hui Ma, and Dieyan Chen, "Afterpulsing and its correction in fluorescence correlation spectroscopy experiments," Appl. Opt. 42, 4031-4036 (2003)

Sort:  Author  |  Year  |  Journal  |  Reset  


  1. D. Madge, E. Elson, W. W. Webb, “Thermodynamic fluctuations in a reacting system measurement by fluorescence correlation spectroscopy,” Phys. Rev. Lett. 29, 705–708 (1972). [CrossRef]
  2. E. L. Elson, D. Madge, “Fluorescence correlation spectroscopy. I. Conceptual basis and theory,” Biopolymers 13, 1–27 (1974). [CrossRef]
  3. D. Madge, E. L. Elson, W. W. Webb, “Fluorescence correlation spectroscopy. II. An experimental realization,” Biopolymers 13, 29–61 (1974). [CrossRef]
  4. R. Rigler, U. Mets, J. Widengren, P. Kask, “Fluorescence correlation spectroscopy with high count rate and low background: analysis of translational diffusion,” Eur. Biophys. J. 22, 169–175 (1993). [CrossRef]
  5. D. E. Koppel, D. Axelrod, J. Schlessinger, E. L. Elson, W. W. Webb, “Dynamics of fluorescence marker concentration as a probe of mobility,” Biophys. J. 16, 1315–1329 (1976). [CrossRef] [PubMed]
  6. H. Qian, E. L. Elson, C. Frieden, “Studies on the structure of actin gels using time correlation spectroscopy of fluorescent beads,” Biophys. J. 63, 1000–1010 (1992). [CrossRef] [PubMed]
  7. P. F. Fahey, D. E. Koppel, L. S. Barak, D. E. Wolf, E. L. Elson, W. W. Webb, “Lateral diffusion in planar lipid bilayers,” Science 195, 305–306 (1977). [CrossRef] [PubMed]
  8. D. Madge, “Chemical kinetics and fluorescence correlation spectroscopy,” Q. Rev. Biophys. 9, 35–47 (1976). [CrossRef] [PubMed]
  9. M. Ehrenberg, R. Rigler, “Rotational Brownian motion and fluorescence intensity fluctuations,” Chem. Phys. 4, 390–401 (1974). [CrossRef]
  10. A. G. I. Palmer, N. L. Thompson, “Molecular aggregation characterized by high order autocorrelation in fluorescence correlation spectroscopy,” Biophys. J. 52, 257–270 (1987). [CrossRef] [PubMed]
  11. H. Qian, E. L. Elson, “Distribution of molecular aggregation by analysis of fluctuation moments,” Proc. Natl. Acad. Sci. USA 87, 5479–5483 (1990). [CrossRef] [PubMed]
  12. J. Widengren, U. Mets, R. Rigler, “Fluorescence correlation spectroscopy of triplet states in solution: a theoretical and experimental study,” J. Phys. Chem. 99, 13,368–13,379 (1995). [CrossRef]
  13. P. Cluzel, M. Surette, S. Leibler, “An ultrasensitive bacterial motor revealed by monitoring signaling proteins in single cells,” Science 287, 1652–1655 (2000). [CrossRef] [PubMed]
  14. E. Overbeck, C. Sinn, “Silicon avalanche photodiodes as detectors for photon correlation experiments,” Rev. Sci. Instrum. 69, 3515–3523 (1998). [CrossRef]
  15. H. C. Burstyn, J. V. Sengers, “Time dependence of critical concentration fluctuations in a binary liquid,” Phys. Rev. A 27, 1071–1085 (1983). [CrossRef]
  16. J. X. Zhu, D. J. Durian, J. Müller, D. A. Weitz, D. J. Pine, “Scaling of transient hydrodynamic interactions in concentrated suspensions,” Phys. Rev. Lett. 68, 2559–2562 (1992). [CrossRef] [PubMed]
  17. M. Hobel, J. Ricka, “Dead-time and afterpulsing correction in multiphoton timing with nonideal detectors,” Rev. Sci. Instrum. 65, 2326–2336 (1994). [CrossRef]
  18. H. C. Burstyn, “Afterpulsing effects in photon correlation experiments,” Rev. Sci. Instrum. 51, 1431–1433 (1980). [CrossRef]

Cited By

Alert me when this paper is cited

OSA is able to provide readers links to articles that cite this paper by participating in CrossRef's Cited-By Linking service. CrossRef includes content from more than 3000 publishers and societies. In addition to listing OSA journal articles that cite this paper, citing articles from other participating publishers will also be listed.

« Previous Article  |  Next Article »

OSA is a member of CrossRef.

CrossCheck Deposited