Summary
Many ion channels, particularly potassium channels, are regulated by intracellular substances, such as nucleotides or Ca2+. These modulators are washed out of the cell during standard whole-cell patch clamp recordings, or maintained at a particular concentration if they are included in the pipette solution. Perforated patch clamp recording permits electrical access between the cell and the patch pipette using pore-forming antibiotics such as nystatin or amphotericin B. These are permeable to small monovalent ions but present a physical barrier to the larger impermeable ions and molecules. This maintains the integrity of many cytoplasmic components including soluble second messengers, and also helps to prevent channel “run down”.
Similar content being viewed by others
References
Horn R. and Marty A. (1988) Muscarinic activation of ionic currents measured by a new whole-cell recording method. J. Gen. Physiol. 2, 145–59.
Rae J., Cooper K., Gates P., and Watsky M. (1991) Low access resistance perforated patch recordings using amphotericin B. J. Neurosci. Methods 37, 15–26.
Horn R. (1991) Diffusion of nystatin in plasma membrane is inhibited by a glass-membrane seal. Biophys. J. 60, 329–33.
Toye A.A., Lippiat J.D., Proks P., Shi-momura K., Bentley L., et al. (2005) A genetic and physiological study of impaired glucose homeostasis control in C57BL/6J mice. Diabetologia 48, 675–86.
Tsuboi T., Lippiat J.D., Ashcroft F.M., and Rutter GA. (2004) ATP-dependent interaction of the cytosolic domains of the inwardly rectifying K+ channel Kir6.2 revealed by fluorescence resonance energy transfer. Proc. Natl Acad. Sci. USA 101, 76–81.
Acknowledgments
I thank Dr. Peter Proks (Oxford) and Dr. John Linley (Leeds) for helpful discussion regarding this technique, and the Welcome Trust and BBSRC for financial support.
Author information
Authors and Affiliations
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2008 Springer Science+Business Media, LLC
About this protocol
Cite this protocol
Lippiat, J.D. (2008). Whole-Cell Recording Using the Perforated Patch Clamp Technique. In: Lippiat, J.D. (eds) Potassium Channels. Methods in Molecular Biology, vol 491. Humana Press. https://doi.org/10.1007/978-1-59745-526-8_11
Download citation
DOI: https://doi.org/10.1007/978-1-59745-526-8_11
Publisher Name: Humana Press
Print ISBN: 978-1-934115-65-7
Online ISBN: 978-1-59745-526-8
eBook Packages: Springer Protocols