Pennell Thomas, Suchyna Thomas, Wang Jianbin, Heo Jinseok, Felske James D, Sachs Frederick, Hua Susan Z
Department of Mechanical and Aerospace Engineering, State University of New York-Buffalo, Buffalo, New York 14260, USA.
Anal Chem. 2008 Apr 1;80(7):2447-51. doi: 10.1021/ac702169t. Epub 2008 Feb 27.
We developed a microfluidic chip that provides rapid temperature changes and accurate temperature control of the perfusing solution to facilitate patch-clamp studies. The device consists of a fluid channel connected to an accessible reservoir for cell culture and patch-clamp measurements. A thin-film platinum heater was placed in the flow channel to generate rapid temperature change, and the temperature was monitored using a thin-film resistor. We constructed the thermal chip using SU-8 on a glass wafer to minimize the heat loss. The chip is capable of increasing the solution temperature from bath temperature (20 degrees C) to 80 degrees C at an optimum heating rate of 0.5 degrees C/ms. To demonstrate the ability of the thermal chip, we have conducted on-chip patch-clamp recordings of temperature-sensitive ion channels (TRPV1) transfected HEK293 cells. The heat-stimulated currents were observed using whole-cell and cell-attached patch configurations. The results demonstrated that the chip can provide rapid temperature jumps at the resolution of single-ion channels.
我们开发了一种微流控芯片,该芯片能使灌注溶液实现快速的温度变化并进行精确的温度控制,以促进膜片钳研究。该装置由一个与用于细胞培养和膜片钳测量的可及储液器相连的流体通道组成。在流动通道中放置了一个薄膜铂加热器以产生快速的温度变化,并使用薄膜电阻器监测温度。我们在玻璃晶圆上使用SU-8构建了热芯片,以尽量减少热损失。该芯片能够以0.5℃/ms的最佳加热速率将溶液温度从浴温(20℃)提高到80℃。为了证明热芯片的能力,我们对转染了温度敏感离子通道(TRPV1)的HEK293细胞进行了芯片上的膜片钳记录。使用全细胞和细胞贴附式膜片配置观察热刺激电流。结果表明,该芯片能够在单离子通道分辨率下提供快速的温度跃升。