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用于电场驱动单细胞捕获与激活的微流控装置。

Microfluidic device for electric field-driven single-cell capture and activation.

作者信息

Toriello Nicholas M, Douglas Erik S, Mathies Richard A

机构信息

Department of Chemistry and UC Berkeley/UCSF Joint Graduate Group in Bioengineering, University of California, Berkeley, California 94720, USA.

出版信息

Anal Chem. 2005 Nov 1;77(21):6935-41. doi: 10.1021/ac051032d.

Abstract

A microchip that performs directed capture and chemical activation of surface-modified single cells has been developed. The cell capture system is comprised of interdigitated gold electrodes microfabricated on a glass substrate within PDMS channels. The cell surface is labeled with thiol functional groups using endogenous RGD receptors, and adhesion to exposed gold pads on the electrodes is directed by applying a driving electric potential. Multiple cell types can thus be sequentially and selectively captured on desired electrodes. Single-cell capture efficiency is optimized by varying the duration of field application. Maximum single-cell capture is attained for the 10-min trial, with 63 +/- 9% (n = 30) of the electrode pad rows having a single cell. In activation studies, single M1WT3 CHO cells loaded with the calcium-sensitive dye fluo-4 AM were captured; exposure to the muscarinic agonist carbachol increased the fluorescence to 220 +/- 74% (n = 79) of the original intensity. These results demonstrate the ability to direct the adhesion of selected living single cells on electrodes in a microfluidic device and to analyze their response to chemical stimuli.

摘要

一种能够对表面修饰的单细胞进行定向捕获和化学激活的微芯片已经研制成功。细胞捕获系统由微制造在聚二甲基硅氧烷(PDMS)通道内玻璃基板上的叉指式金电极组成。利用内源性RGD受体,细胞表面用硫醇官能团进行标记,通过施加驱动电势引导细胞与电极上暴露的金垫粘附。因此,多种细胞类型可以在所需电极上依次、选择性地捕获。通过改变电场施加时间来优化单细胞捕获效率。在10分钟的试验中实现了最大单细胞捕获量,63±9%(n = 30)的电极垫行有单个细胞。在激活研究中,捕获了加载钙敏染料fluo-4 AM的单个M1WT3 CHO细胞;暴露于毒蕈碱激动剂卡巴胆碱后,荧光增加到原始强度的220±74%(n = 79)。这些结果证明了在微流控装置中引导选定活单细胞在电极上粘附并分析其对化学刺激反应的能力。

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