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硅纳米线和光刺激在细胞内和细胞间电耦合研究中的应用。

Silicon Nanowires and Optical Stimulation for Investigations of Intra- and Intercellular Electrical Coupling.

机构信息

The James Franck Institute, The University of Chicago;

Department of Chemistry, The University of Chicago.

出版信息

J Vis Exp. 2021 Jan 28(167). doi: 10.3791/61581.

Abstract

Myofibroblasts can spontaneously internalize silicon nanowires (SiNWs), making them an attractive target for bioelectronic applications. These cell-silicon hybrids offer leadless optical modulation capabilities with minimal perturbation to normal cell behavior. The optical capabilities are obtained by the photothermal and photoelectric properties of SiNWs. These hybrids can be harvested using standard tissue culture techniques and then applied to different biological scenarios. We demonstrate here how these hybrids can be used to study the electrical coupling of cardiac cells and compare how myofibroblasts couple to one another or to cardiomyocytes. This process can be accomplished without special equipment beyond a fluorescent microscope with coupled laser line. Also shown is the use of a custom-built MATLAB routine that allows the quantification of calcium propagation within and between the different cells in the culture. Myofibroblasts are shown to have a slower electrical response than that of cardiomyocytes. Moreover, the myofibroblast intercellular propagation shows slightly slower, though comparable velocities to their intracellular velocities, suggesting passive propagation through gap junctions or nanotubes. This technique is highly adaptable and can be easily applied to other cellular arenas, for in vitro as well as in vivo or ex vivo investigations.

摘要

肌成纤维细胞可以自发内化硅纳米线(SiNWs),使它们成为生物电子应用的有吸引力的目标。这些细胞-硅的混合物具有无铅光学调制能力,对正常细胞行为的干扰最小。光学性能是通过 SiNWs 的光热和光电特性获得的。这些混合物可以使用标准的组织培养技术收获,然后应用于不同的生物学场景。我们在这里展示了这些混合物如何用于研究心脏细胞的电偶联,并比较肌成纤维细胞彼此之间或与心肌细胞之间的偶联方式。这个过程可以在配备耦合激光线的荧光显微镜之外的普通设备下完成。还展示了使用自定义 MATLAB 例程来定量测量培养物中不同细胞内和细胞间钙的传播。结果表明,肌成纤维细胞的电响应比心肌细胞慢。此外,肌成纤维细胞的细胞间传播速度虽然稍慢,但与细胞内传播速度相当,这表明通过缝隙连接或纳米管进行被动传播。该技术具有高度的适应性,可以很容易地应用于其他细胞领域,包括体外、体内和离体研究。

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