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The OpenPicoAmp-100k: an open-source high-performance amplifier for single channel recording in planar lipid bilayers

  • Vadim ShlyonskyEmail author
  • David Gall
Ion channels, receptors and transporters
  • 65 Downloads
Part of the following topical collections:
  1. Ion channels, receptors and transporters

Abstract

We propose an upgraded version of our previously designed open-source lipid bilayer amplifier. This improved amplifier is now suitable both for the use in introductory courses in biophysics and neurosciences at the undergraduate level and for scientific research. Similar to its predecessor, the OpenPicoAmp-100k is designed using the common lithographic printed circuit board fabrication process and off-the-shelf electronic components. It consists of the high-speed headstage, followed by voltage-gain amplifier with built-in 6-order Bessel filter. The amplifier has a bandwidth of 100 kHz in the presence of 100-pF input membrane capacitance and is capable of measuring ion channel current with amplitudes from sub-pA and up to ± 4 nA. At the full bandwidth and with a 1 GΩ transimpedance gain, the amplifier shows 12 pArms noise with an open input and 112 pArms noise in the presence of 100-pF input capacitance, while at the 5-kHz bandwidth (typical in single-channel experiments), noise amounts to 0.45 pArms and 2.11 pArms, respectively. Using an optocoupler circuit producing TTL-controlled current impulses and using 50% threshold analysis, we show that at full bandwidth, the amplifier has deadtimes of 3.5 μs and 5 μs at signal-to-noise ratios (SNR) of 9 and 1.7, respectively. Near 100% of true current impulses longer than 5 μs and 6.6 μs are detected at these two respective SNRs, while false event detection rate remains acceptably low. The wide bandwidth of the amplifier was confirmed in bilayer experiments with alamethicin, for which open ion channel current events shorter that 10 μs could be resolved.

Keywords

Voltage-clamp Temporal resolution Ion channels Electronic design 

Notes

Acknowledgments

We gratefully acknowledge Freddy Dupuis for his technical help in the design of the rail splitter and Prof. Fabrice Homblé for the critical reading of the manuscript.

Funding information

This project has received support from the Fonds d’Encouragement à l’Enseignement of the Université libre de Bruxelles. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.

Supplementary material

424_2019_2319_MOESM1_ESM.docx (1.8 mb)
ESM 1 (DOCX 1792 kb)

References

  1. 1.
    Averbuch AZ, Eisenberg RS, Israeli M, Schuss Z (1994) Detecting ionic currents in single channels using wavelet analysis, part I: zero mean Gaussian noise. SPIE Proc 2303:76–90.  https://doi.org/10.1117/12.188812 CrossRefGoogle Scholar
  2. 2.
    Baaken G, Sondermann M, Schlemmer C, Rühe J, Behrends JC (2008) Planar microelectrode-cavity array for high-resolution and parallel electrical recording of membrane ionic currents. Lab Chip 8:938–944CrossRefGoogle Scholar
  3. 3.
    Bean RC, Shepherd WC, Chan H, Eichner J (1969) Discrete conductance fluctuations in lipid bilayer protein membranes. J Physiol 53:741–757Google Scholar
  4. 4.
    Bertrand D, Bader CR, Distasi C, Forster IC (1989) Single-channel current simulation and recording using a photodiode as current generator. J Neurosci Methods 26:233–238CrossRefGoogle Scholar
  5. 5.
    Cole KS (1949) Dynamic electrical characteristics of the squid axon membrane. Arch Sci Physiol 3:253–258Google Scholar
  6. 6.
    Colquhoun D, Sigworth FJ (1995) Fitting and Statistical Analysis of Single-Channel Records. In: Sakmann B, Neher E (eds) Single-Channel Recording. Springer, BostonGoogle Scholar
  7. 7.
    Costa JA, Nguyen DA, Leal-Pinto E, Gordon RE, Hanss B (2013) Wicking: a rapid method for manually inserting ion channels into planar lipid bilayers. PLoS One 8:e60836CrossRefGoogle Scholar
  8. 8.
    Draguhn A, Pfeiffer M, Heinemann U, Polder R (1997) A simple hardware model for the direct observation of voltage-clamp performance under realistic conditions. J Neurosci Methods 78:105–113CrossRefGoogle Scholar
  9. 9.
    Hamill OP, Marty A, Neher E, Sakmann B, Sigworth FJ (1981) Improved patch-clamp techniques for high-resolution current recording from cells and cell-free membrane patches. Pflugers Arch 391:85–100CrossRefGoogle Scholar
  10. 10.
    Hartel AJW, Shekar S, Ong P, Schroeder I, Thiel G, Shepard KL (2019) High bandwidth approaches in nanopore and ion channel recordings - A tutorial review. Anal Chim Acta 1061:13–27CrossRefGoogle Scholar
  11. 11.
    Hladky SB, Haydon DA (1970) Discreteness of conductance change in bimolecular lipid membranes in the presence of certain antibiotics. Nature 225:451–453CrossRefGoogle Scholar
  12. 12.
    Marmont G (1949) Studies on the axon membrane. I A new method. J Cell Comp Physiol 34:351–382CrossRefGoogle Scholar
  13. 13.
    Mayer M, Kriebel JK, Tosteson MT, Whitesides GM (2003) Microfabricated teflon membranes for low-noise recordings of ion channels in planar lipid bilayers. Biophys J 85:2684–2695CrossRefGoogle Scholar
  14. 14.
    Pein F, Tecuapetla-Gomez I, Schutte OM, Steinem C, Munk A (2018) Fully automatic multiresolution idealization for filtered ion channel recordings: flickering event detection. IEEE Trans Nanobiosci 17:300–320CrossRefGoogle Scholar
  15. 15.
    Rosenstein JK, Ramakrishnan S, Roseman J, Shepard KL (2013) Single ion channel recordings with CMOS-anchored lipid membranes. Nano Lett 13:2682–2686CrossRefGoogle Scholar
  16. 16.
    Rouzrokh A, Ebrahimi SA, Mahmoudian M (2009) Construction, calibration, and validation of a simple patch-clamp amplifier for physiology education. Adv Physiol Educ 33:121–129CrossRefGoogle Scholar
  17. 17.
    Schibel AE, Edwards T, Kawano R, Lan W, White HS (2010) Quartz nanopore membranes for suspended bilayer ion channel recordings. Anal Chem 82:7259–7266CrossRefGoogle Scholar
  18. 18.
    Shah SI, Demuro A, Mak DD, Parker I, Pearson JE, Ullah G (2018) TraceSpecks: a software for automated idealization of noisy patch-clamp and imaging data. Biophys J 115:9–21CrossRefGoogle Scholar
  19. 19.
    Shapovalov G, Lester HA (2004) Gating transitions in bacterial ion channels measured at 3 microsecond resolution. J Gen Physiol 124:151–161CrossRefGoogle Scholar
  20. 20.
    Shekar S, Chien CC, Hartel A, Ong P, Clarke OB, Marks A, Drndic M, Shepard KL (2019) Wavelet denoising of high-bandwidth nanopore and ion-channel signals. Nano Lett 19:1090–1097CrossRefGoogle Scholar
  21. 21.
    Shlyonsky V, Dupuis F, Gall D (2014) The OpenPicoAmp: an open-source planar lipid bilayer amplifier for hands-on learning of neuroscience. PLoS One 9:e108097CrossRefGoogle Scholar
  22. 22.
    Sigworth FJ (1995) Electronic design of the patch clamp. In: Sakmann B, Neher E (eds) Single-Channel Recording. Springer, BostonGoogle Scholar
  23. 23.
    Silverman BW (1999) Wavelets in statistics: Beyond the standard assumptions. Phil Trans R Soc A 357:2459–2473CrossRefGoogle Scholar
  24. 24.
    The Axon guide, electrophysiology and biophysics laboratory techniques. Molecular Devices, SunnyvaleGoogle Scholar
  25. 25.
    Wonderlin WF, Finkel A, French RJ (1990) Optimizing planar lipid bilayer single-channel recordings for high resolution with rapid voltage steps. Biophys J 58:289–297CrossRefGoogle Scholar
  26. 26.
    Zhang B, Galusha J, Shiozawa PG, Wang G, Bergren AJ, Jones RM, White RJ, Ervin EN, Cauley CC, White HS (2007) Bench-top method for fabricating glass-sealed nanodisk electrodes, glass nanopore electrodes, and glass nanopore membranes of controlled size. Anal Chem 79:4778–4787CrossRefGoogle Scholar

Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2019

Authors and Affiliations

  1. 1.Laboratoire de Physiologie et Pharmacologie, Faculté de MédecineUniversité libre de BruxellesBrusselsBelgium
  2. 2.Laboratoire d’Enseignement de la Physique, Faculté de MédecineUniversité libre de BruxellesBrusselsBelgium

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