A. Alrifaiy, N. Bitaraf, M. Druzin, O. Lindahl and K. Ramser
We present a novel approach to reproduce hypoxia on a chip by patch-clamp investigations on single nerve cells exposed to anoxic and normoxic environments. The patch-clamp technique was combined with microfluidics to enable fast exchange of buffer-solutions. The micropipette was included within the microfluidic chip to allow investigations with full control over the oxygen content in vicinity of the sample. Nerve cells from Sprague Dawley rats were prepared and inserted into the channels of the microchip. Single nerve cells were optically trapped and manipulated to be patched by patch-clamp micropipette. The experiments were aimed to test proof of principle and to perform patchclamp electrophysiological measurements on the cells under well-defined conditions. The oxygen level within the microfluidic channels was in the range of 0.5 to 1.5%. The laser tweezers showed no remarkable photo-induced effect on the investigated nerve cells and no effects on the electrophysiological measurements were detected. The approach of using closed microfluidic system in patch-clamp experiments showed significant advantages to control the oxygen concentration around the investigated cell. This may be adapted to be used in other biological investigations of single cells demanding optimal control of the surroundings.
DOI
We present a novel approach to reproduce hypoxia on a chip by patch-clamp investigations on single nerve cells exposed to anoxic and normoxic environments. The patch-clamp technique was combined with microfluidics to enable fast exchange of buffer-solutions. The micropipette was included within the microfluidic chip to allow investigations with full control over the oxygen content in vicinity of the sample. Nerve cells from Sprague Dawley rats were prepared and inserted into the channels of the microchip. Single nerve cells were optically trapped and manipulated to be patched by patch-clamp micropipette. The experiments were aimed to test proof of principle and to perform patchclamp electrophysiological measurements on the cells under well-defined conditions. The oxygen level within the microfluidic channels was in the range of 0.5 to 1.5%. The laser tweezers showed no remarkable photo-induced effect on the investigated nerve cells and no effects on the electrophysiological measurements were detected. The approach of using closed microfluidic system in patch-clamp experiments showed significant advantages to control the oxygen concentration around the investigated cell. This may be adapted to be used in other biological investigations of single cells demanding optimal control of the surroundings.
DOI
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