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使用蛋白质功能化孔进行细胞表征。

Cell characterization using a protein-functionalized pore.

作者信息

Carbonaro Andrea, Mohanty Swomitra K, Huang Haiyan, Godley Lucy A, Sohn Lydia L

机构信息

Dept. of Mechanical Engineering, University of California at Berkeley, Berkeley, CA 94720-1740, USA.

出版信息

Lab Chip. 2008 Sep;8(9):1478-85. doi: 10.1039/b801929k. Epub 2008 Jul 25.

Abstract

We demonstrate a highly-sensitive and label-free method for characterizing cells based on cell-surface receptors. The method involves measuring a current pulse generated when an individual cell passes through an artificial pore. When the pore is functionalized with proteins, specific interactions between a cell-surface marker and the functionalized proteins retard the cell, thus leading to an increased pulse duration that indicates the presence of that specific biomarker. For proof-of-principle, we successfully screened murine erythroleukemia cells based on their CD34 surface marker in both a single and mixed population of cells. Further, we developed a unified constrained statistical model for estimating the ratios of cells in a mixed population. Finally, we demonstrated our ability to screen a small number of cells (hundreds or less) with high accuracy and sensitivity. Overall, our pore-based method is broadly applicable and, in the future, could provide a full range of in vitro cell-based assays.

摘要

我们展示了一种基于细胞表面受体来表征细胞的高灵敏度且无标记的方法。该方法涉及测量单个细胞通过人工孔时产生的电流脉冲。当孔用蛋白质功能化时,细胞表面标志物与功能化蛋白质之间的特异性相互作用会使细胞减速,从而导致脉冲持续时间增加,这表明存在该特定生物标志物。为了验证原理,我们成功地在单个细胞群体和混合细胞群体中基于其CD34表面标志物对小鼠红白血病细胞进行了筛选。此外,我们开发了一个统一的约束统计模型来估计混合细胞群体中细胞的比例。最后,我们展示了以高精度和高灵敏度筛选少量细胞(数百个或更少)的能力。总体而言,我们基于孔的方法具有广泛的适用性,并且在未来可以提供一系列基于体外细胞的检测方法。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/61c4/12345329/2f586b1c09d1/nihms-2094721-f0001.jpg

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