Abstract
Solutions used in patch-clamp can be divided into two general categories by the side of the patch membrane with which the solution is in contact. The compositions of extracellular solutions are usually similar to those of the natural extracellular fluids, whereas intracellular solutions resemble cytosol with respect to pH, free Ca2+, and Mg2+ concentrations and ATP content. Designing solutions for patch-clamp experiments requires careful consideration of the physicochemical processes controlling the concentrations of protons and divalent cations, ionic strength, and osmotic pressure. Liquid junction potentials arising at the interfaces between different experimental solutions may contribute to the membrane potential and should be measured or calculated and adequately compensated. This chapter describes general strategies and methods for practical calculations of these parameters, which are critically important for successful patch-clamp experiments.
This is a preview of subscription content, log in via an institution to check access.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
Similar content being viewed by others
References
Prosser CL (1973) Inorganic ions. In: Ladd Prosser C (ed) Comparative animal physiology. Saunders, Philadelphia, pp 79–110
Sabirov RZ, Okada Y, Oiki S (1997) Two-sided action of protons on an inward rectifier K+ channel (IRK1). Pflugers Arch 433:428–434
Sabirov RZ, Prenen J, Droogmans G, Nilius B (2000) Extra- and intracellular proton-binding sites of volume-regulated anion channels. J Membr Biol 177:13–221
Tsai TD, Shuck ME, Thompson DP, Bienkowski MJ, Lee KS (1995) Intracellular H+ inhibits a cloned rat kidney outer medulla K+ channel expressed in Xenopus oocytes. Am J Physiol 268:C1173–C1178
Robinson RA, Stokes RH (1959) Electrolyte solutions. Butterworths, London
Miller DJ, Smith GL (1984) EGTA purity and the buffering of calcium ions in physiological solutions. Am J Physiol 246:C160–C166
Oiki S, Yamamoto T, Okada Y (1994) Apparent stability constants and purity of Ca-chelating agents evaluated using Ca-selective electrodes by the double-log optimization method. Cell Calcium 15:209–216
Oiki S, Yamamoto T, Okada Y (1994) A simultaneous evaluation method of purity and apparent stability constant of Ca-chelating agents and selectivity coefficient of Ca- selective electrodes. Cell Calcium 15:199–208
Harrison SM, Bers DM (1987) The effect of temperature and ionic strength on the apparent Ca- affinity of EGTA and the analogous Ca-chelators BAPTA and dibromo-BAPTA. Biochim Biophys Acta 925:133–143
Harrison SM, Bers DM (1989) Correction of proton and Ca association constants of EGTA for temperature and ionic strength. Am J Physiol 256:C1250–C1256
Smith GL, Miller DJ (1985) Potentiometric measurements of stoichiometric and apparent affinity constants of EGTA for protons and divalent ions including calcium. Biochim Biophys Acta 839:287–299
Fabiato A (1988) Computer programs for calculating total from specified free or free from specified total ionic concentrations in aqueous solutions containing multiple metals and ligands. Methods Enzymol 157:378–417
Neher E (1992) Correction for liquid junction potentials in patch clamp experiments. Methods Enzymol 207:123–131
D’Ans J, Surawsky H, Synowietz C (1977) In: Schafer Kl (ed) Densities of liquid systems and their heat capacities, Landolt-Bornstein numerical data and functional relationships in science and technology. New Series. Springer, Berlin
Barry PH, Lynch JW (1991) Liquid junction potentials and small cell effects in patch-clamp analysis. J Membr Biol 121:101–117
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2012 Springer
About this protocol
Cite this protocol
Sabirov, R.Z., Morishima, S. (2012). Solutions for Patch-Clamp Experiments. In: Okada, Y. (eds) Patch Clamp Techniques. Springer Protocols Handbooks. Springer, Tokyo. https://doi.org/10.1007/978-4-431-53993-3_28
Download citation
DOI: https://doi.org/10.1007/978-4-431-53993-3_28
Publisher Name: Springer, Tokyo
Print ISBN: 978-4-431-53992-6
Online ISBN: 978-4-431-53993-3
eBook Packages: Springer Protocols