Time-correlated single photon counting continues to gain importance in a wide range of applications. Most prominently, it is used for time-resolved fluorescence measurements with sensitivity down to the single molecule level. While the primary goal of the method used to be the determination of fluorescence lifetimes upon optical excitation by short light pulses, recent modifications and refinements of instrumentation and methodology allow for the recovery of much more information from the detected photons, and enable entirely new applications. This is achieved most successfully by continuously recording individually detected photons with their arrival time and detection channel information (time tagging), thus avoiding premature data reduction and concomitant loss of information. An important property of the instrumentation used is the number of detection channels and the way they interrelate. Here we present a new instrument architecture that allows scalability in terms of the number of input channels while all channels are synchronized to picoseconds of relative timing and yet operate independent of each other. This is achieved by means of a modular design with independent crystal-locked time digitizers and a central processing unit for sorting and processing of the timing data. The modules communicate through high speed serial links supporting the full throughput rate of the time digitizers. Event processing is implemented in programmable logic, permitting classical histogramming, as well as time tagging of individual photons and their temporally ordered streaming to the host computer. Based on the time-ordered event data, any algorithms and methods for the analysis of fluorescence dynamics can be implemented not only in postprocessing but also in real time. Results from recently emerging single molecule applications are presented to demonstrate the capabilities of the instrument.
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December 2008
Research Article|
December 29 2008
Scalable time-correlated photon counting system with multiple independent input channels
Michael Wahl;
1
PicoQuant GmbH
, Rudower Chaussee 29, D-12489 Berlin, Germany
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Hans-Jürgen Rahn;
Hans-Jürgen Rahn
1
PicoQuant GmbH
, Rudower Chaussee 29, D-12489 Berlin, Germany
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Tino Röhlicke;
Tino Röhlicke
1
PicoQuant GmbH
, Rudower Chaussee 29, D-12489 Berlin, Germany
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Gerald Kell;
Gerald Kell
2
Fachhochschule Brandenburg
, Magdeburger Str. 50, D-14770 Brandenburg, Germany
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Daniel Nettels;
Daniel Nettels
3Biochemisches Institut,
Universität Zürich
, Winterthurerstrasse 190, CH-8057 Zürich, Switzerland
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Frank Hillger;
Frank Hillger
3Biochemisches Institut,
Universität Zürich
, Winterthurerstrasse 190, CH-8057 Zürich, Switzerland
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Ben Schuler;
Ben Schuler
3Biochemisches Institut,
Universität Zürich
, Winterthurerstrasse 190, CH-8057 Zürich, Switzerland
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Rainer Erdmann
Rainer Erdmann
1
PicoQuant GmbH
, Rudower Chaussee 29, D-12489 Berlin, Germany
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a)
Author to whom correspondence should be addressed. Electronic mail: wahl@picoquant.com.
Rev. Sci. Instrum. 79, 123113 (2008)
Article history
Received:
July 11 2008
Accepted:
December 03 2008
Citation
Michael Wahl, Hans-Jürgen Rahn, Tino Röhlicke, Gerald Kell, Daniel Nettels, Frank Hillger, Ben Schuler, Rainer Erdmann; Scalable time-correlated photon counting system with multiple independent input channels. Rev. Sci. Instrum. 1 December 2008; 79 (12): 123113. https://doi.org/10.1063/1.3055912
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