The authors wish to thank the EPSRC, BBSRC and MRC who funded this work as part of the IRC in Bionanotechnology. Support from British Heart Foundation funding to G.L.S. is acknowledged.
University of Heidelberg (2006) Proc Physiol Soc 4, PC11
Poster Communications: Norbert Klauke2, Jon M. Cooper2, Godfrey Smith1
1. Biomedical and Life Sciences, Glasgow University, Glasgow, United Kingdom. 2. Department of Electronics, Glasgow University, Glasgow, United Kingdom.
Biophysical studies of isolated cardiac myocytes would benefit from devices that allow the rapid microfluidic manipulation of the extracellular space around limited regions of a single cell. This abstract describes a method to partition the extracellular space around a single cardiac myocyte bridging a hydrophobic gap, produced with polymer soft-lithography. This system allows the two ends of a single cardiac cell to be independently superfused using an arrangement of concentric micropipettes. The inner pipette is used for delivery of the superfusate and the outer pipette for the removal of the extracellular solution around the cell ends. An additional concentric pipette system, loaded with a separate test solution (containing drugs, ions or permeabilisation reagents) can be inserted into a selected compartment for variable periods using a piezo-stepper, enabling digitally controlled rapid solution switching within the picolitre volume around either end of the cell. Planar gold electrodes were also integrated into the device, enabling electrical stimulation and recording of the evoked action potential. The device was mounted on an inverted microscope to allow sarcomere length and epifluorescence measurements. This system allows the demonstration of spatially restricted effects of the test solution by observing the response at one end of the cardiac cell in comparison with the untreated region. Initial measurements using this novel device include: (i) the regional permeabilization of the surface membrane of the cardiac cell and the subsequent perfusion of the intracellular space; (ii) the local application of caffeine thereby generating spatially confined intracellular Ca2+ transients; (iii) differential recording across the high resistance (~50MΩ) between the separate compartments to reveal the extracellular signal associated with an evoked action potential.
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Where applicable, experiments conform with Society ethical requirements.