Single-molecule-sensitive microscopy and spectroscopy are transforming biophysics and materials science laboratories. Techniques such as fluorescence correlation spectroscopy (FCS) and single-molecule sensitive fluorescence resonance energy transfer (FRET) are now commonly available in research laboratories but are as yet infrequently available in teaching laboratories. We describe inexpensive electronics and open-source software that bridges this gap, making state-of-the-art research capabilities accessible to undergraduates interested in biophysics. We include a discussion of the intensity correlation function relevant to FCS and how it can be determined from photon arrival times. We demonstrate the system with a measurement of the hydrodynamic radius of a protein using FCS that is suitable for the undergraduate teaching laboratory. The FPGA-based electronics, which are easy to construct, are suitable for more advanced measurements as well, and several applications are described. As implemented, the system has 8 ns timing resolution, can control up to four laser sources, and can collect information from as many as four photon-counting detectors.
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July 2014
APPARATUS AND DEMONSTRATION NOTES|
July 01 2014
Inexpensive electronics and software for photon statistics and correlation spectroscopy
Benjamin D. Gamari;
Benjamin D. Gamari
Department of Physics, University of Massachusetts
, Amherst, Massachusetts 01002
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Dianwen Zhang;
Dianwen Zhang
a)
Department of Physics, University of Massachusetts
, Amherst, Massachusetts 01002
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Richard E. Buckman;
Richard E. Buckman
Department of Physics, University of Massachusetts
, Amherst, Massachusetts 01002
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Peker Milas;
Peker Milas
Department of Physics, University of Massachusetts
, Amherst, Massachusetts 01002
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John S. Denker;
John S. Denker
AT&T Laboratories Research (ret.), Florham Park NJ, 07932
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Hui Chen;
Hui Chen
Wadsworth Center, New York State Department of Health
, 120 New Scotland Ave, Albany, New York 12208
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Hongmin Li;
Hongmin Li
Wadsworth Center, New York State Department of Health
, 120 New Scotland Ave, Albany, New York 12208 and Department of Biomedical Sciences, School of Public Health, State University of New York at Albany
, P.O. BOX 509, Albany, New York 12201-0509
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Lori S. Goldner
Lori S. Goldner
b)
Department of Physics, University of Massachusetts
, Amherst, Massachusetts 01002
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a)
Present address: Beckman Institute for Advanced Science and Technology, University of Illinois at Urbana-Champaign, 405 North Mathews, Urbana, Illinois 61801.
b)
Author to whom correspondence should be addressed. Electronic mail: [email protected]
Am. J. Phys. 82, 712–722 (2014)
Article history
Received:
June 14 2012
Accepted:
March 10 2014
Citation
Benjamin D. Gamari, Dianwen Zhang, Richard E. Buckman, Peker Milas, John S. Denker, Hui Chen, Hongmin Li, Lori S. Goldner; Inexpensive electronics and software for photon statistics and correlation spectroscopy. Am. J. Phys. 1 July 2014; 82 (7): 712–722. https://doi.org/10.1119/1.4869188
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