Pflügers Archiv

, Volume 412, Issue 3, pp 322–327 | Cite as

A temperature and solution control system for the measurement of single channel currents in excised membrane patches

  • J. W. Lynch
  • P. H. Barry
  • N. Quartararo
Excitable Tissues and Central Nervous Physiology Instruments and Techniques


A technique is described which permits accurate temperature control and relatively rapid temperature changes (within about 2 min for 10°C changes between 10° and 40°C) of the solution perfusing the exposed surface of excised membrane patches. The simultaneous exchange of temperature controlled solution is also possible. Using the “sleeve technique”, patches excised from cells in standard tissue culture dishes are removed to a separate chamber where temperature and solution are accurately controlled. This avoids two common limitations of existing temperature or solution control systems: (1) test solution contamination of the tissue perfusion solution which may impair cell viability and (2) the use of specialised chambers which are unsuitable for use with cultured cells. In the system described, temperature control is possible over the range of at least 4–40°C. Desired temperatures can be preset to within approximately ±1°C, and can then be controlled and measured to an accuracy of ±0.1°C. At a constant temperature, the system enables rapid solution changes, the solution bathing the excised patch being exchanged in approximately 3s.

Key words

Single channel currents Temperature Solution Excised membrane patches 


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  1. Barrett JN, Magleby KL, Pallotta BS (1982) Properties of single calcium-activated potassium channels in cultured rat muscle. J Physiol (Lond) 331:211–230Google Scholar
  2. Brett RS, Dilger JP, Adams PR, Lancaster B (1986) A method for the rapid exchange of solutions bathing excised membrane patches. Biophys J 50:987–992Google Scholar
  3. Cannell MB, Lederer WJ (1986) A novel experimental chamber for single-cell voltage-clamp and pathch-clamp applications with low electrical noise and excellent temperature and flow control. Pflügers Arch 406:536–539Google Scholar
  4. Cull-Candy SG, Parker I (1983) Experimental approaches to examine single glutamate-receptor ion channels in locust muscle fibres. In: Sakman B, Neher E (eds) Single-channel recurding. Plenum Press, New York, pp 389–400Google Scholar
  5. Datyner NB, Gintant GA, Cohen IS (1985) Versatile temperature controlled tissue bath for studies of isolated cells using an inverted microscope. Pflügers Arch 403:318–323Google Scholar
  6. Fenwick WM, Marty A, Neher E (1982) A patch-clamp study of bovine chromaffin cells and of their sensitivity to acetylcholine. J Physiol (Lond) 331:577–597Google Scholar
  7. Franke C, Hatt H, Dudel J (1987) Liquid filament switch for ultrafast exchange of solutions at excised patches of synaptic membrane of crayfish muscle. Neurosci Lett 77:199–204Google Scholar
  8. Hamill OP, Marty A, Neher E, Sakmann B, Sigworth FJ (1981) Improved patch-clamp techniques for high-resolution current from cells and cell-free membrane patches. Pflügers Arch 391:85–100Google Scholar
  9. Hering S, Beech DJ, Bolton TB (1987) A simple method of fast extracellular solution exchange for the study of whole-cell or single channel currents using the patch-clamp technique. Pflügers Arch 410:335–337Google Scholar
  10. Kakei M, Ashcroft FM (1987) A microflow superfusion system for use with excised membrane patches. Pflügers Arch 409:337–341Google Scholar
  11. Konigsberg IR (1979) Skeletal myoblasts in culture. In: Jakoby WB, Pastan IH (eds) Methods in enzymology, vol lvii: cell culture. Academic Press, New York, pp 511–527Google Scholar
  12. Lee KS, Akaike N, Brown AM (1980) The suction pipette method for internal perfusion and voltage clamp of small excitable cells. J Neurosci Methods 2:51–78Google Scholar
  13. Marty A, Neher E (1983) Tight-seal whole-cell recording. In: Sakmann B, Neher E (eds) Single-channel recording. Plenum Press, New York, pp 107–122Google Scholar
  14. Rae JL, Levis RA (1984) Patch voltage clamp of lens epithelial cells: theory and practice. Mol Physiol 6:115–162Google Scholar
  15. Wuartararo N, Barry PH (1987) A simple technique for transferring excised patches of membrane to different solutions for single channel measurements. Pflügers Arch 410:677–678Google Scholar
  16. Yellen G (1982) Single Ca2+-activated non-selective cation channels in neuroblastoma cells. Nature 296:357–359Google Scholar

Copyright information

© Springer-Verlag 1988

Authors and Affiliations

  • J. W. Lynch
    • 1
  • P. H. Barry
    • 1
  • N. Quartararo
    • 1
  1. 1.School of Physiology and PharmacologyUniversity of New South WalesKensingtonAustralia

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