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Review of Scientific Instruments / Browse Journal / Volume 79 / Issue 12 / OPTICS; ATOMS AND MOLECULES; SPECTROSCOPY; PHOTON DETECTORS

Rev. Sci. Instrum. 79, 123113 (2008); doi:10.1063/1.3055912 (8 pages)

Scalable time-correlated photon counting system with multiple independent input channels

Michael Wahl1, Hans-Jürgen Rahn1, Tino Röhlicke1, Gerald Kell2, Daniel Nettels3, Frank Hillger3, Ben Schuler3, and Rainer Erdmann1

1PicoQuant GmbH, Rudower Chaussee 29, D-12489 Berlin, Germany Map This map
2Fachhochschule Brandenburg, Magdeburger Str. 50, D-14770 Brandenburg, Germany Map This map
3Biochemisches Institut, Universität Zürich, Winterthurerstrasse 190, CH-8057 Zürich, Switzerland Map This map

(Received 11 July 2008; accepted 3 December 2008; published online 29 December 2008)

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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.

© 2008 American Institute of Physics

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KEYWORDS and PACS

Keywords

analogue-digital conversion, fluorescence, molecule-photon collisions, photodetectors, photoexcitation, photon counting, radiative lifetimes, time resolved spectra

PACS

  • 85.60.Gz

    Photodetectors (including infrared and CCD detectors)

  • 07.57.-c

    Infrared, submillimeter wave, microwave and radiowave instruments and equipment

  • 33.50.Dq

    Fluorescence and phosphorescence spectra

  • 33.70.Ca

    Oscillator and band strengths, lifetimes, transition moments, and Franck-Condon factors

  • 33.80.-b

    Photon interactions with molecules

ARTICLE DATA

Permalink
http://link.aip.org/link/RSINAK/v79/i12/p123113/s1

PUBLICATION DATA

ISSN:

0034-6748 (print)  
1089-7623 (online)

Publisher:

$abstract.society.value is a Member of CrossRef American Institute of Physics

  1. J. R. Lakowicz, Principles of Fluorescence Spectroscopy (Springer, New York, 1999).
  2. J. Kalisz, Metrologia 41, 17 (2004). [Inspec] [ISI]
  3. R. Rigler and E. S. Elson, Fluorescence Correlation Spectroscopy Theory and Applications (Springer, Berlin, 2001).
  4. K. Schätzel, Inst. Phys. Conf. Ser. 77, 175-184 (1985).
  5. M. Ehrenberg and R. Rigler, Chem. Phys. 4, 390 (1974). [Inspec] [ISI]
  6. P. Kask, P. Piksarv, and U. Mets, Eur. Biophys. J. 12, 163 (1985). [Inspec] [ISI]
  7. T. Basche, W. E. Moerner, M. Orrit, and H. Talon, Phys. Rev. Lett. 69, 1516 (1992). [MEDLINE]
  8. P. Tinnefeld, K. D. Weston, T. Vosch, M. Cotlet, T. Weil, J. Hofkens, K. Mullen, F. C. De Schryver, and M. Sauer, J. Am. Chem. Soc. 124, 14310 (2002). [ISI] [MEDLINE]
  9. J. Sykora, K. Kaiser, I. Gregor, W. Bonigk, G. Schmalzing, and J. Enderlein, Anal. Chem. 79, 4040 (2007). [MEDLINE]
  10. S. Fore, T. A. Laurence, C. W. Hollars, and T. Huser, IEEE J. Sel. Top. Quantum Electron. 13, 996 (2007).
  11. P. Kask, P. Piksarv, U. Mets, M. Pooga, and E. Lippmaa, Eur. Biophys. J. 14, 257 (1987). [Inspec] [MEDLINE]
  12. A. J. Berglund, A. C. Doherty, and H. Mabuchi, Phys. Rev. Lett. 89, 068101 (2002). [ISI] [MEDLINE]
  13. D. Nettels, I. V. Gopich, A. Hoffmann, and B. Schuler, Proc. Natl. Acad. Sci. U.S.A. 104, 2655 (2007). [MEDLINE]
  14. C. Eggeling, J. R. Fries, L. Brand, R. Gunther, and C. A. M. Seidel, Proc. Natl. Acad. Sci. U.S.A. 95, 1556 (1998). [ISI] [MEDLINE]
  15. M. Wahl, R. Erdmann, K. Lauritsen, and H. J. Rahn, Proc. SPIE 3259, 173 (1998).
  16. J. B. Edel and A. J. Demello, Appl. Phys. Lett. 90, 053904 (2007)APPLAB000090000005053904000001.
  17. H. Z. Lin, S. R. Tabaei, D. Thomsson, O. Mirzov, P. O. Larsson, and I. G. Scheblykin, J. Am. Chem. Soc. 130, 7042 (2008). [MEDLINE]
  18. C. Eggeling, S. Berger, L. Brand, J. R. Fries, J. Schaffer, A. Volkmer, and C. A. M. Seidel, J. Biotechnol. 86, 163 (2001). [ISI] [MEDLINE]
  19. H. P. Lu, Methods in Molecular Biology (Humana Press, Totowa, NJ, 2005), Vol. 305, p. 385.
  20. M. Wahl, I. Gregor, M. Patting, and J. Enderlein, Opt. Express 11, 3583 (2003). [SPIN] [ISI]
  21. D. Nettels and B. Schuler, IEEE J. Sel. Top. Quantum Electron. 13, 990 (2007).
  22. D. C. Lamb, A. Schenk, C. Rocker, C. Scalfi-Happ, and G. U. Nienhaus, Biophys. J. 79, 1129 (2000). [ISI] [MEDLINE]
  23. M. Böhmer, M. Wahl, H. J. Rahn, R. Erdmann, and J. Enderlein, Chem. Phys. Lett. 353, 439 (2002). [Inspec] [ISI]
  24. F. Koberling, M. Wahl, M. Patting, H. J. Rahn, P. Kapusta, and R. Erdmann, Proc. SPIE 5143, 181 (2003).
  25. U. Ortmann, T. Dertinger, M. Wahl, H. J. Rahn, M. Patting, and R. Erdmann, Proc. SPIE 5325, 179 (2004).
  26. S. Felekyan, R. Kuhnemuth, V. Kudryavtsev, C. Sandhagen, W. Becker, and C. A. M. Seidel, Rev. Sci. Instrum. 76, 083104 (2005)RSINAK000076000008083104000001. [ISI]
  27. M. Wahl, H. J. Rahn, I. Gregor, R. Erdmann, and J. Enderlein, Rev. Sci. Instrum. 78, 033106 (2007)RSINAK000078000003033106000001. [ISI] [MEDLINE]
  28. M. Ghioni, A. Gulinatti, I. Rech, F. Zappa, and S. Cova, IEEE J. Sel. Top. Quantum Electron. 13, 852 (2007).
  29. I. Rech, D. Resnati, S. Marangonia, M. Ghioni, and S. Cova, Proc. SPIE 6771, 77113 (2007).
  30. I. Rech, S. Marangonia, D. Resnati, M. Ghioni, and S. Cova, J. Mod. Opt. 99999(1), 1–8 (2008), http://www.informaworld.com/10.1080/09500340802318309. Accessed Dec. 18, 2008.
  31. D. J. S. Birch, D. McLoskey, A. Sanderson, K. Suhling, and A. S. Holmes, J. Fluoresc. 4, 91 (1994).
  32. K. D. Weston, M. Dyck, P. Tinnefeld, C. Muller, D. P. Herten, and M. Sauer, Anal. Chem. 74, 5342 (2002). [ISI] [MEDLINE]
  33. D. P. Herten, A. Biebricher, M. Heilemann, T. Heinlein, C. Müller, P. Schlüter, P. Tinnefeld, K. D. Weston, M. Sauer, and J. Wolfrum, Recent Res. Dev. Appl. Phys. 7, 345 (2004).
  34. H. Ta, A. Mogk, B. Bukau, and D. -P. Herten, 14th International Workshop on Single Molecule Spectroscopy and Ultrasensitive Analysis, Berlin, 2008 (unpublished).
  35. W. Becker, A. Bergmann, C. Biskup, L. Kelbauskas, T. Zimmer, N. Klöcker, and K. Benndorf, Proc. SPIE 4963, 175 (2003).
  36. D. Knoll, K. E. Ehwald, B. Heinemann, A. Fox, K. Blum, H. Rucker, F. Furnhammer, B. Senapati, R. Barth, and U. Haak, Tech. Dig. - Int. Electron Devices Meet. 2002, 783.
  37. F. Hillger, D. Hänni, D. Nettels, S. Geister, M. Grandin, M. Textor, and B. Schuler, Angew. Chem., Int. Ed. 47, 6184 (2008). [MEDLINE]
  38. F. U. Hartl and M. Hayer-Hartl, Science 295, 1852 (2002). [MEDLINE]
  39. R. H. Brown and R. Q. Twiss, Nature (London) 177, 27 (1956).
  40. F. Hillger, D. Nettels, S. Dorsch, and B. Schuler, J. Fluoresc. 17, 759 (2007). [MEDLINE]
  41. P. Kask, P. Piksarv, M. Pooga, U. Mets, and E. Lippmaa, Biophys. J. 55, 213 (1989). [Inspec]
  42. B. Schuler, E. A. Lipman, P. J. Steinbach, M. Kumke, and W. A. Eaton, Proc. Natl. Acad. Sci. U.S.A. 102, 2754 (2005). [MEDLINE]
  43. A. A. Deniz, T. A. Laurence, M. Dahan, D. S. Chemla, P. G. Schultz, and S. Weiss, Annu. Rev. Phys. Chem. 52, 233 (2001). [MEDLINE]
  44. For getting more accurate estimates for the transfer efficiency, it would be necessary to correct for background photons, for different quantum yields of Alexa 488 and Alexa 594, and for the different detection efficiencies and crosstalk between the detection channels, B. Schuler, Methods Mol. Biol. 350, 115 (2007) (for the mere discrimination between subpopulations, these corrections can be ignored). [MEDLINE]
  45. R. Best, K. Merchant, I. V. Gopich, B. Schuler, A. Bax, and W. A. Eaton, Proc. Natl. Acad. Sci. U.S.A. 104, 18964 (2007). [MEDLINE]
  46. S. Weiss, Science 283, 1676 (1999). [MEDLINE]
  47. G. Haran, J. Phys.: Condens. Matter 15, R1291 (2003). [ISI]
  48. X. Michalet, S. Weiss, and M. Jäger, Chem. Rev. (Washington, D.C.) 106, 1785 (2006). [MEDLINE]
  49. B. Schuler and W. A. Eaton, Curr. Opin. Struct. Biol. 18, 16 (2008). [MEDLINE]

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Figures (4) Tables (1)

Figures (click on thumbnails to view enlargements)

FIG. 1
Block diagram of the instrument architecture. Dots between identical building blocks denote scalability.
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FIG. 2
Results of intrinsic performance measurements: DNL (flat line) and timing jitter in four channels (peaked histograms).
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FIG. 3
FCS curves of 500 pM GroEL-bound rhodanese labeled with Alexa 488. The autocorrelation G∥ ∥(τ) of the detected parallel polarized photons is shown in black, and the cross correlation G∥⊥(τ) between parallel and perpendicularly polarized photons in gray. (a) Logarithmic plot, illustrating the large difference in amplitude from rotational dynamics, but coincidence of the correlations for τ>1 μs. (b) Linear plot for τ<1 μs.
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FIG. 4
Subpopulation-resolved FCS of a mixture of 100 pM chaperone-bound rhodanese, labeled with Alexa 488, and 200 pM of Pro20, labeled with a FRET dye pair (donor, Alexa 488 and acceptor, Alexa 594). (a) Transfer efficiency histogram with two peaks corresponding to the two subpopulations. The ranges of Eapp used for extracting the autocorrelation functions [(b) and (c)] of horizontally polarized donor emission G∥ ∥(τ) are shaded correspondingly. (b) G∥ ∥(τ) for the subpopulation of chaperone-bound rhodanese. (c) G∥ ∥(τ) for the Pro20 subpopulation.
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Tables

Table I. Long span timing accuracy and jitter for different time spans within and across channels (all in nanoseconds).

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