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Optics Express

Optics Express

  • Editor: Andrew M. Weiner
  • Vol. 21, Iss. 20 — Oct. 7, 2013
  • pp: 23543–23555

High countrate real-time FCS using F2Cor

Emmanuel Schaub  »View Author Affiliations


Optics Express, Vol. 21, Issue 20, pp. 23543-23555 (2013)
http://dx.doi.org/10.1364/OE.21.023543


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Abstract

We present a fluorescence correlation spectroscopy setup based on a software correlator. The setup can process autocorrelation curves in real-time at countrate as high as 8MHz, with time resolution of 1µs. It uses the F2Cor autocorrelation algorithm, a low cost counting board and a desktop computer. Symmetrical normalization, which improves the signal to noise ratio of the FCS curve for large values of the lag-time, is adapted to the F2Cor algorithm. A new acquisition mode, which we call oscilloscope-mode, is presented. It takes advantage of the flexibility F2Cor, and proves to be very useful for optical setup adjustment. As an application of this setup, we performed FCS measurements on a reference tetramethylrhodamine solution at high concentration, up to 2.5µM, which extend to the micromolar range the concentration applicable in FCS, using a conventional optical setup. At such high countrates the FCS curves need to be corrected for dead-time of the photo-detector, which was done successfully.

© 2013 Optical Society of America

OCIS Codes
(070.4550) Fourier optics and signal processing : Correlators
(120.6200) Instrumentation, measurement, and metrology : Spectrometers and spectroscopic instrumentation
(170.2520) Medical optics and biotechnology : Fluorescence microscopy
(070.7145) Fourier optics and signal processing : Ultrafast processing

ToC Category:
Spectroscopy

History
Original Manuscript: July 16, 2013
Revised Manuscript: September 13, 2013
Manuscript Accepted: September 19, 2013
Published: September 26, 2013

Citation
Emmanuel Schaub, "High countrate real-time FCS using F2Cor," Opt. Express 21, 23543-23555 (2013)
http://www.opticsinfobase.org/oe/abstract.cfm?URI=oe-21-20-23543


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