A microfluidic device to confine a single cardiac myocyte in a sub-nanoliter volume on planar microelectrodes for extracellular potential recordings.
A hybrid chip is described which combines a microfluidic network fabricated in a silicone elastomer (PDMS) with planar microelectrodes. It was used to measure extracellular potentials from single adult murine cardiac myocytes in a restricted extracellular space. The recorded variations in the extracellular potentials were caused by transmembrane currents associated with spontaneously initiated intracellular calcium waves. Single cells were trapped inside the 100 pl microchamber by pressure gradients and maintained for several hours by continuous perfusion. In addition, the localized delivery of drugs to a portion of the cell was demonstrated. The impedance of the electrodes was reduced by a factor of 10 to 20 after the electrodeposition of platinum black. Biopotentials recorded from single cells with platinum black electrodes showed a three-fold decrease in the noise, resulting in a maximum signal-to-noise ratio of 15:1. Characteristic variations in the frequency and shape of the extracellular potentials were observed among different cells which are most likely due to differences in the sarcoplasmic reticulum Ca(2+) load. Our device architecture permits the integration of electrochemical and optical sensors for multiparameter recordings.
Department of Biomedical Engineering, Vanderbilt University, Nashville, TN, USA.
Andreas A
AA
Werdich
Eduardo A
EA
Lima
Borislav
B
Ivanov
Igor
I
Ges
Mark E
ME
Anderson
John P
JP
Wikswo
Franz J
FJ
Baudenbacher
D000818:Animals; D004129:Dimethylpolysiloxanes / Q000737:chemistry; D005110:Extracellular Space / Q000737:chemistry / Q000502:physiology*; D008564:Membrane Potentials / Q000502:physiology*; D051379:Mice; D008839:Microelectrodes; D044085:Microfluidics / Q000295:instrumentation / Q000379:methods*; D032383:Myocytes, Cardiac / Q000166:cytology* / Q000502:physiology*; D036103:Nanotechnology / Q000379:methods*; D011108:Polymers / Q000737:chemistry; D012828:Silicones / Q000737:chemistry
D004129:Dimethylpolysiloxanes; D011108:Polymers; D012828:Silicones