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A first step towards practical single cell proteomics: a microfluidic antibody capture chip with TIRF detection

机译:迈向实用单细胞蛋白质组学的第一步:具有TIRF检测功能的微流控抗体捕获芯片

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摘要

We have developed a generic platform to undertake the analysis of protein copy number from single cells. The approach described here is 'all-optical' whereby single cells are manipulated into separate analysis chambers using an optical trap; single cells are lysed by a shock wave caused by laser-induced microcavitation, and the protein released from a single cell is measured by total internal reflection microscopy as it is bound to micro-printed antibody spots within the device. The platform was tested using GFP transfected cells and the relative precision of the measurement method was determined to be 88%. Single cell measurements were also made on a breast cancer cell line to measure the relative levels of unlabelled human tumour suppressor protein p53 using a chip incorporating an antibody sandwich assay format. These results suggest that this is a viable method for measuring relative protein levels in single cells.
机译:我们已经开发了一个通用平台来进行单细胞蛋白质拷贝数的分析。这里描述的方法是“全光学”的,其中单个细胞使用一个光阱被操纵到单独的分析室中。激光诱导的空化作用引起的冲击波裂解了单细胞,并且由于单细胞释放的蛋白质与设备内的微印抗体斑点结合,因此通过全内反射显微镜对单细胞释放的蛋白质进行了测量。使用GFP转染的细胞对平台进行测试,测定方法的相对精度确定为88%。还使用包含抗体夹心测定形式的芯片在乳腺癌细胞系上进行了单细胞测量,以测量未标记的人类肿瘤抑制蛋白p53的相对水平。这些结果表明,这是一种测量单个细胞中相对蛋白水平的可行方法。

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