Microfluidic partitioning of the extracellular space around single cardiac myocytes.
This paper describes the partitioning of the extracellular space around an electrically activated single cardiac myocyte, constrained within a microfluidic device. Central to this new method is the production of a hydrophobic gap-structure, which divides the extracellular space into two distinct microfluidic pools. The content of these pools was controlled using a pair of concentric automated pipets (subsequently called "dual superfusion pipet"), each providing the ability to dispense (i.e., the source, inner pipet) and aspirate (the sink, outer pipet) a buffer solution (perfusate) into each of the two pools. For rapid solution switching around the cell, additional dual superfusion pipets were inserted into the microchannel for defined time periods using a piezostepper, enabling us to add a test solution, such as a drug. Three distinct areas of the cell were manipulated, namely, the microfluidic environment, the cellular membrane, and the intracellular space. Planar integrated microelectrodes enabled the electrical stimulation of the cardiomyocyte and the recording of the evoked action potential. The device was mounted on an inverted microscope to allow simultaneous sarcomere length and epifluorescence measurements during evoked electrical activity, including, for example, the response of the stimulated end of the cardiac myocyte in comparison with the untreated cell end.
Klauke
Norbert
N
Department of Electronics, University of Glasgow, Glasgow G12 8LT, Scotland. nobert@elec.gla.ac.uk
Smith
Godfrey L
GL
Cooper
Jonathan M
JM
D000200:Action Potentials; D000818:Animals; D002021:Buffers; D002462:Cell Membrane; D005110:Extracellular Space; D005453:Fluorescence; D006801:Humans; D008839:Microelectrodes; D046210:Microfluidic Analytical Techniques; D044085:Microfluidics / Q000379:methods*; D032383:Myocytes, Cardiac / Q000166:cytology*; D010477:Perfusion; D012518:Sarcomeres
D002021:Buffers