Skip to main content

State-of-the-Art Automated Patch Clamp: Heat Activation, Action Potentials, and High Throughput in Ion Channel Screening

  • Protocol
  • First Online:
Patch-Clamp Methods and Protocols

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1183))

Abstract

A successful robotic approach of the patch clamp technique is based on planar patch clamp chips where a glass pipette, as used in conventional patch clamping, is replaced by a thin planar glass sheet with a small hole in the middle. Automated patch clamp (APC) systems utilizing this chip design offer higher throughput capabilities and ease of use and thus have become common in basic research, drug development, and safety screening. Further development of existing devices and introduction of new systems widen the range of possible experiments and increase throughput. Here, two features with different areas of applications that meet the needs of drug discovery researchers and basic researchers alike are described. The utilized system is a medium throughput APC device capable of recording up to eight cells simultaneously. The temperature control capability and the possibility to perform recordings not only in the voltage clamp but also in the current clamp mode are described in detail. Since eight recordings can be generated in parallel without compromising data quality, reliable and cost-effective and time-effective screening of compounds against ion channels using voltage clamp and current clamp electrophysiology can be performed.

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

Access this chapter

Protocol
USD 49.95
Price excludes VAT (USA)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
eBook
USD 99.00
Price excludes VAT (USA)
  • Available as EPUB and PDF
  • Read on any device
  • Instant download
  • Own it forever
Softcover Book
USD 129.99
Price excludes VAT (USA)
  • Compact, lightweight edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info
Hardcover Book
USD 199.99
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

References

  1. Neher E, Sakmann B (1976) Single channel currents recorded from membrane of denervated frog muscle fibres. Nature 260:799–802

    Article  CAS  PubMed  Google Scholar 

  2. Dunlop J, Bowlby M, Peri R et al (2008) High-throughput electrophysiology: an emerging paradigm for ion-channel screening and physiology. Nat Rev 7:358–368

    CAS  Google Scholar 

  3. Farre C, Haythornthwaite A, Haarmann C et al (2009) Port-a-patch and patchliner: high fidelity electrophysiology for secondary screening and safety pharmacology. Comb Chem High Throughput Screen 12:24–37

    Article  CAS  PubMed  Google Scholar 

  4. Stoelzle S, Obergrussberger A, Brüggemann A et al (2011) State-of-the art automated patch-clamp devices: heat activation, action potentials, and high throughput in ion channel screening. Front Pharmacol 2:1–11

    Article  Google Scholar 

  5. Polonchuk L (2009) Toward a new gold standard for early safety: automated temperature-controlled hERG test on the Patchliner®. Front Pharmacol 3:3

    Google Scholar 

  6. Milligan CJ, Li J, Sukumar P, Majeed Y et al (2009) Robotic multiwell planar patch-clamp for native and primary mammalian cells. Nat Protoc 4:244–255

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  7. Balansa W, Islam R, Fontaine F et al (2010) Ircinialactams: subunit-selective glycine receptor modulators from Australian sponges of the family Irciniidae. Bioorg Med Chem 18:2912–2919

    Article  CAS  PubMed  Google Scholar 

  8. Stoelzle S, Haythornthwaite A, Kettenhofen R et al (2011) Automated patch-clamp on mESC-derived cardiomyocytes for cardiotoxicity prediction. J Biomol Screen 16:910–916

    Article  CAS  PubMed  Google Scholar 

  9. Farre C, Stoelzle S, Haarman C et al (2007) Automated ion channel screening: patch-clamping made easy. Expert Opin Ther Targets 11:557–565

    Article  CAS  PubMed  Google Scholar 

  10. Brüggemann A, Farre C, Haarmann C, Haythornthwaite A et al (2008) Planar patch-clamp: advances in electrophysiology. Methods Mol Biol 491:165–176

    Article  PubMed  Google Scholar 

  11. Majeed Y, Bahnasi Y, Seymour V et al (2010) Rapid and contrasting effects of rosiglitazone on transient receptor potential TRPM3 and TRPC5 channels. Mol Pharmacol 79:1023–1030

    Article  Google Scholar 

  12. Peier A, Reeve AJ, Andersson DA et al (2002) A heat-sensitive TRP channel expressed in keratinocytes. Science 296:2046–2049

    Article  CAS  PubMed  Google Scholar 

  13. Xu H, Ramsey IS, Kotecha SA et al (2002) TRPV3 is a calcium-permeable temperature-sensitive cation channel. Nature 418:181–186

    Article  CAS  PubMed  Google Scholar 

  14. Smith GD, Gunthorpe MJ, Kelsell RE et al (2002) TRPV3 is a temperature-sensitive vanilloid receptor-like protein. Nature 418:186–190

    Article  CAS  PubMed  Google Scholar 

  15. Chung MK, Lee H, Mizuno A et al (2004) 2-aminoethoxydiphenyl borate activates and sensitizes the heat-gated ion channel TRPV3. J Neurosci 24:5177–5182

    Article  CAS  PubMed  Google Scholar 

  16. Kolossov E, Bostani T, Roell W et al (2006) Engraftment of engineered ES cell-derived cardiomyocytes but not BM cells restores contractile Function to the infarcted myocardium. J Exp Med 203:2315–2327

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  17. Hu H, Grandl J, Bandell M et al (2009) Two amino acid residues determine 2-APB sensitivity of the ion channels TRPV3 and TRPV4. Proc Natl Acad Sci U S A 106:1626–1631

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  18. Becker N, Stoelzle S, Göpel S et al (2013) Minimized cell usage for stem cell-derived and primary cells on an automated patchclamp system. J Pharmacol Toxicol Meth 68(1):82–87, pii: S1056-8719(13)00232-3

    Article  CAS  Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to Sonja Stoelzle-Feix .

Editor information

Editors and Affiliations

Rights and permissions

Reprints and permissions

Copyright information

© 2014 Springer Science+Business Media New York

About this protocol

Cite this protocol

Stoelzle-Feix, S. (2014). State-of-the-Art Automated Patch Clamp: Heat Activation, Action Potentials, and High Throughput in Ion Channel Screening. In: Martina, M., Taverna, S. (eds) Patch-Clamp Methods and Protocols. Methods in Molecular Biology, vol 1183. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-1096-0_4

Download citation

  • DOI: https://doi.org/10.1007/978-1-4939-1096-0_4

  • Published:

  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-1095-3

  • Online ISBN: 978-1-4939-1096-0

  • eBook Packages: Springer Protocols

Publish with us

Policies and ethics