Skip to main content
SpringerLink
Log in

Modern TCSPC Electronics: Principles and Acquisition Modes

  • Chapter
  • First Online:
Advanced Photon Counting

Part of the book series: Springer Series on Fluorescence ((SS FLUOR,volume 15))

Abstract

Time-correlated single-photon counting (TCSPC) is an extraordinarily versatile and sensitive technique. While it was initially used almost only to measure excited state lifetimes, it can today be used much more flexibly, embracing and combining experimental methods that in the past required separate instrumentation. This has become possible by time-tagged event recording and modern time measurement circuitry. This chapter shows how such technologies operate with regard to electronics, data processing, and applications. Some implementation details will be exemplified by state-of-the-art TCSPC instruments and a recent software package for TCSPC data acquisition and analysis.

This is a preview of subscription content, log in via an institution to check access.

Access this chapter

Log in via an institution
Chapter
USD 29.95
Price excludes VAT (USA)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
eBook
USD 349.00
Price excludes VAT (USA)
  • Available as EPUB and PDF
  • Read on any device
  • Instant download
  • Own it forever
Hardcover Book
USD 449.99
Price excludes VAT (USA)
  • Durable hardcover edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info

Tax calculation will be finalised at checkout

Purchases are for personal use only

Institutional subscriptions

Notes

  1. 1.

    This naming convention appears a little arbitrary because it has evolved historically. See the related footnote on T3 mode for the original historical meaning.

  2. 2.

    As outlined in the footnote on T2 mode, this nomenclature is somewhat arbitrary and only explicable in historical context. The abbreviation T3 stems from T3R which in turn stands for time-tagged-time-resolved (TTTR). This was essentially an ad hoc term to adequately describe our first implementation of early time-tagged TCSPC by an extension of the classical stopwatch scheme by a lower-resolution time tag [27]. The idea was originally conceived for the purpose of single molecule detection in capillary flow [28] but had not been widely recognized then. Other early implementers of the concept (Becker & Hickl GmbH, unpublished at the time) and related publications [29] referred to it only from a specialized application or implementation perspective, using the terms burst-integrated fluorescence lifetime (BIFL) or FIFO mode. Today it is more common to speak of TTTR or time tagging as the overall method with T2 and T3 modes as its variants. T3 mode was called T3 mode because it is close to the historical T3R scheme.

References

  1. Bollinger LM, Thomas GE (1961) Measurement of the time dependence of scintillation intensity by a delayed coincidence method. Rev Sci Instrum 32:1044–1050

    Article  CAS  Google Scholar 

  2. Connor DVO, Phillips D (1984) Time-correlated single photon counting. Academic Press, London

    Google Scholar 

  3. Lakowicz JR (2006) Principles of fluorescence spectroscopy, 3rd edn. Springer Science + Business Media, New York

    Book  Google Scholar 

  4. Bülter A (2014) Single-photon counting detectors for the visible range between 300 nm and 1000 nm. In: Kapusta P et al. (eds) Advanced photon counting: applications, methods, instrumentation. Springer series on fluorescence. Springer International Publishing, doi: 10.1007/4243_2014_63

  5. Buller GS, Collins RJ (2014) Single-photon detectors for infrared wavelengths in the range 1 to 1.7 μm. In: Kapusta P et al. (eds) Advanced photon counting: applications, methods, instrumentation. Springer series on fluorescence. Springer International Publishing, doi: 10.1007/4243_2014_64

  6. Becker W (2005) Advanced time-correlated single photon counting techniques. Springer, Berlin

    Book  Google Scholar 

  7. Rossi B, Nereson N (1946) Experimental arrangement for the measurement of small time intervals between the discharges of Geiger-Müller counters. Rev Sci Instrum 17:65–71

    Article  CAS  Google Scholar 

  8. Kalisz J (2004) Review of methods for time interval measurements with picosecond resolution. Metrologia 41:17–32

    Article  Google Scholar 

  9. Roberts GW, Ali-Bakhshian M (2010) A brief introduction to time-to-digital and digital-to-time converters. IEEE Transact Circ Syst II Expr Briefs 57:153–157

    Article  Google Scholar 

  10. Henzler S (2010) Time-to-digital converters. Springer, Dordrecht/Heidelberg/London/New York

    Book  Google Scholar 

  11. Heinemann B et al. (2010) SiGe HBT technology with fT/fmax of 300 GHz/500 GHz and 2.0 ps CML gate delay. Technical digest, IEEE international electron device meeting (IEDM), San Francisco, 06–08 Dec 2010, pp 688–691

    Google Scholar 

  12. Wahl M, Röhlicke T, Rahn HJ, Erdmann R, Kell G, Ahlrichs A, Kernbach M, Schell AW, Benson O (2013) Integrated multichannel photon timing instrument with very short dead time and high throughput. Rev Sci Instrum 084:043102

    Article  Google Scholar 

  13. Elson E, Magde D (1974) Fluorescence correlation spectroscopy. I conceptual basis and theory. Biopolymers 13:1–27

    Article  CAS  Google Scholar 

  14. Thompson NL, Lieto AM, Allen NW (2002) Recent advances in fluorescence correlation spectroscopy. Curr Opin Struc Biol 12:634–641

    Article  Google Scholar 

  15. Dertinger T, Rüttinger S (2014) Advanced FCS: an introduction to fluorescence lifetime correlation spectroscopy and dual-focus FCS. In: Kapusta P et al. (eds) Advanced photon counting: applications, methods, instrumentation. Springer series on fluorescence. Springer International Publishing, doi: 10.1007/4243_2014_72

  16. Böhmer M, Wahl M, Rahn HJ, Erdmann R, Enderlein J (2002) Time-resolved fluorescence correlation spectroscopy. Chem Phys Lett 353:439–445

    Article  Google Scholar 

  17. Enderlein J, Gregor I (2005) Using fluorescence lifetime for discriminating detector after-pulsing in fluorescence-correlation spectroscopy. Rev Sci Instrum 76:033102

    Article  Google Scholar 

  18. Felekyan S, Kalinin S, Valeri A, Seidel CAM (2009) Filtered FCS and species cross correlation function. In: Periasamy A, So PTC (eds) Multiphoton microscopy in the biomedical sciences IX; Proceedings of SPIE 7183:71830D:1–71830D:12

    Google Scholar 

  19. Felekyan S, Kalinin S, Sanabria H, Valeri A, Seidel CAM (2012) Filtered FCS: species auto- and cross-correlation functions highlight binding and dynamics in biomolecules. Chem Phys Chem 13:1036–1053

    CAS  Google Scholar 

  20. Eggeling C, Berger S, Brand L, Fries JR, Schaffer J, Volkmer A, Seidel CAM (2001) Data registration and selective single-molecule analysis using multi-parameter fluorescence detection. J Biotechnol 86:163–180

    Article  CAS  Google Scholar 

  21. Ishii K, Tahara T (2010) Resolving inhomogeneity using lifetime-weighted fluorescence correlation spectroscopy. J Phys Chem B 114:12383–12391

    Article  CAS  Google Scholar 

  22. Ishii K, Tahara T (2013) Two-dimensional fluorescence lifetime correlation spectroscopy. J Phys Chem B 117:11414–11432

    Article  CAS  Google Scholar 

  23. Otosu T, Tahara T (2014) Lifetime-weighted FCS and 2D FLCS: advanced application of time-tagged TCSPC. In: Kapusta P et al. (eds) Advanced photon counting: applications, methods, instrumentation. Springer series on fluorescence. Springer International Publishing, doi: 10.1007/4243_2014_65

  24. Grußmayer KS, Herten D-P (2014) Photon Antibunching in Single Molecule Fluorescence Spectroscopy. In: Kapusta P et al. (eds) Advanced photon counting: applications, methods, instrumentation. Springer series on fluorescence. Springer International Publishing, doi: 10.1007/4243_2014_71

  25. Wahl M, Rahn HJ, Röhlicke T, Kell G, Nettels D, Hillger F, Schuler B, Erdmann R (2008) Scalable time-correlated photon counting system with multiple independent input channels. Rev Sci Instrum 79:123113

    Article  Google Scholar 

  26. Birch DSJ, McLoskey D, Sanderson A, Suhling K, Holmes AS (1994) Multiplexed time-correlated single-photon counting. J Fluoresc 04:91–102

    Article  CAS  Google Scholar 

  27. Wahl M, Erdmann R, Lauritsen K, Rahn HJ (1998) Hardware solution for continuous time-resolved burst detection of single molecules in flow. Proc SPIE 3259:173–178

    CAS  Google Scholar 

  28. Wilkerson CW Jr, Goodwin PM, Ambrose WP, Martin JC, Keller RA (1993) Detection and lifetime measurement of single molecules in flowing sample streams by laser-induced fluorescence. Appl Phys Lett 062:2030–2033

    Article  Google Scholar 

  29. Eggeling C, Fries JR, Brand L, Gunther R, Seidel CAM (1998) Monitoring conformational dynamics of a single molecule by selective fluorescence spectroscopy. Proc Natl Acad Sci U S A 95:1556–1561

    Article  CAS  Google Scholar 

  30. Wahl M, Röhlicke T, Rahn HJ, Buschmann V, Bertone N, Kell G (2013) High speed multichannel time-correlated single photon counting electronics based on SiGe integrated time-to-digital converters. Proc SPIE 8727:87270W

    Google Scholar 

  31. LaVision GmbH (2014) Ultra-fast gated cameras. http://www.lavision.de/en/products/cameras/ultrafast_gated_cameras.php. Accessed 3 April 2014

  32. Koberling F, Wahl M, Patting M, Rahn HJ, Kapusta P, Erdmann R (2003) Two channel fluorescence lifetime microscope with two colour laser excitation, single-molecule sensitivity and submicrometer resolution. Proc SPIE 5143:181–192

    CAS  Google Scholar 

  33. Ortmann U, Dertinger T, Wahl M, Rahn HJ, Patting M, Erdmann R (2004) Compact TCSPC upgrade package for laser scanning microscopes based on 375 to 470 nm picosecond diode lasers. Proc SPIE 5325:179–186

    Google Scholar 

  34. Li LQ, Davis LM (1995) Rapid and efficient detection of single chromophore molecules in aqueous solution. Appl Opt 34(18):3208–3217

    Article  CAS  Google Scholar 

  35. Davis LM, Williams PE, Ball DA, Swift KM, Matayoshi ED (2003) Data reduction methods for application of fluorescence correlation spectroscopy to pharmaceutical drug discovery. Curr Pharm Biotechnol 04:451–462

    Article  CAS  Google Scholar 

  36. Schätzel K (1985) New concepts in correlator design. In: Institute of Physics conference series, vol 77. Hilger, London, pp 175–184

    Google Scholar 

  37. Wahl M, Gregor I, Patting M, Enderlein J (2003) Fast calculation of fluorescence correlation data with asynchronous time-correlated single-photon counting. Opt Express 11:03583–03591

    Article  Google Scholar 

  38. Yang H, Xie XS (2002) Probing single molecule dynamics photon by photon. J Chem Phys 117:10965–10979

    Article  CAS  Google Scholar 

  39. Yang H, Luo G, Karnchanaphanurach P, Louie TM, Rech I, Cova S, Xun L, Xie XS (2003) Protein conformational dynamics probed by single-molecule electron transfer. Science 302(5643):262–266

    Article  CAS  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Instrumentation Division, PicoQuant GmbH, Rudower Chaussee 29, 12489, Berlin, Germany

    Michael Wahl

Authors
  1. Michael Wahl
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Michael Wahl .

Editor information

Editors and Affiliations

  1. J. Heyrovsky Institute of Physical Chemistry, Academy of Sciences of the Czech Republic, Prague 8, Czech Republic

    Peter Kapusta

  2. PicoQuant GmbH, Berlin, Germany

    Michael Wahl

  3. PicoQuant GmbH, Berlin, Germany

    Rainer Erdmann

Rights and permissions

Reprints and permissions

Copyright information

© 2014 Springer International Publishing Switzerland

About this chapter

Cite this chapter

Wahl, M. (2014). Modern TCSPC Electronics: Principles and Acquisition Modes. In: Kapusta, P., Wahl, M., Erdmann, R. (eds) Advanced Photon Counting. Springer Series on Fluorescence, vol 15. Springer, Cham. https://doi.org/10.1007/4243_2014_62

Download citation

Publish with us

Policies and ethics

Search

Navigation