Summary
A variant of the whole-cell patch clamp technique is described which allows measurement of whole-cell ionic currents in small cells while minimizing cell dialysis with the pipette solution. The technique involves the application of negative pressure to the inside of small (< 1 µm) tip diameter pipettes placed on the cell surface to achieve high resistance seals and membrane rupture. The technique has been used successfully in a variety of different types of cells to study membrane currents carried by Ca and K, currents generated by exchange carriers as well as electrical coupling between cells. Overall, the technique seems well suited for the study of ionic currents in small cells, and provides an alternative to conventional patch clamping techniques which necessitate intracellular dialysis.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Neher E, Sakmann B: Single-channel currents recorded from membrane of denervated frog muscle fibres. Nature 260:779–802, 1976
Neher E, Sakmann B, Steinbach JH: The extracellular patch clamp: a method of resolving currents through individual open channels in biological membranes. Pflugers Arch 375:219–228, 1978
Hamill, OP, Marty A, Neher E, Sakmann B, Sigworth FJ: Improved patch clamp techniques for high-resolution current recording from cells and cell-free membrane patches. Pflugers Arch 391:85–100, 1981
Sakmann B, Neher E: Patch clamp techniques for studying ionic channels in excitable membranes. Ann Rev Physiol 46:455–472, 1984
Byerly L, Hagiwara S: Calcium currents in internally perfused nerve cell bodies of Lymnaea stagnalis. J Physiol 322:503–528, 1982
Uehara A, Hume JR: Interactions of organic Ca channel antagonists with Ca channels in single frog atrial cells. J Gen Physiol 85:621–647, 1985
Hume JR: Comparative interactions of organic Ca channel antagonists with myocardial Ca and K channels. J Pharmacol Exp Ther 234:134–140, 1985
Hume JR, Uehara A: Properties of ‘creep currents’ in single frog atrial cells. J Gen Physiol 87:833–855, 1986
Hume JR, Uehara A: ‘Creep currents’ in single frog atrial cells may be generated by electrogenic Na/Ca-exchange. J Gen Physiol 87:857–884, 1986
Hume JR, Giles W: Active and passive electrical properties of single bullfrog atrial cells. J Gen Physiol 78:19–42, 1981
Hume JR, Giles W: Ionic currents in single isolated bullfrog atrial cells. J Gen Physiol 81:153–194, 1983
Hume JR, Uehara A: Ionic basis of the different action potential configurations of single guinea-pig atrial and ventricular myocytes. J Physiol 368:525–544, 1985
Lee KS, Akaike N, Brown AM: The suction pipette method for internal perfusion and voltage clamp of small excitable cells. J Neurosci Methods 2:51–78, 1980
Tarr M, Trank JW: Chemically mediated cell-to-cell contractile activation in isolated frog atrial cardiac cells. Experientia 34:1472–1474, 1978
Simmons MA, Creazzo T, Hartzell HC: A time-dependent and voltage-sensitive K current in single cells from frog atrium. J Gen Physiol 88:739–756, 1986
Hume JR, Giles W, Robinson K, Shibata EF, Nathan RD, Kanai K, Rassmusson R: A time- and voltage-dependent K current in single cardiac cells from bullfrog atrium. J Gen Physiol 88:777–798, 1986
de Hemptinne A: Properties of the outward currents in frog atrial muscle. Pflugers Arch 329:321–331, 1971
Brown HF, Clark A, Noble SJ: Analysis of pacemaker and repolarization currents in frog atrial muscle. J Physiology 258:547–577, 1976
Hadley RW, Hume JR: An intrinsic potential-dependent inactivation mechanism associated with calcium channels in guinea-pig myocytes. J Physiol 389:205–222, 1987
Inoue R, Kitamura K, Kuriyama H: Two Ca-dependent K-channels classified by the application of tetraethylammonium distribute to smooth muscle membranes of the rabbit portal vein. Pflugers Arch 405: 173–179
Inoue R, Okabe K, Kitamura K, Kuriyama H: A newly identified Ca dependent K channel in the smooth muscle membrane of single cells dispersed from rabbit portal vein. Pflugers Arch 406:138–143, 1986
Wilde DW, Hume JR: Dual action of minoxidil-sulfate on calcium and potassium currents in smooth muscle cells of rabbit portal vein. Circulation: 76: IV 329, 1987
Lowenstein WR, Kanno K, Socolar SJ: Quantum jumps of conductance during formation of membrane channels at cell-cell junction. Nature 274:133–136, 1978
Spray DC, Bennett MVL: Physiology and pharmacology of gap junctions. Ann Rev Physiol 47:281–303, 1985
Veenstra RD, DeHann RL: Measurement of single channel currents from cardiac gap junctions. Science 233:972–974, 1986
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Hume, J.R., Leblanc, R.N. A whole-cell patch clamp technique which minimizes cell dialysis. Mol Cell Biochem 80, 49–57 (1989). https://doi.org/10.1007/BF00231003
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF00231003