Tunable Single-Cell Extraction for Molecular Analyses
Tunable Single-Cell Extraction for Molecular Analyses
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DOI：
10.1016/j.cell.2016.06.025
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发表时间：
2016-07-14
期刊：
影响因子：
64.5
通讯作者：
Vorholt, Julia A.
中科院分区：
文献类型：
--
作者：
Guillaume-Gentil, Orane;Grindberg, Rashel V.;Vorholt, Julia A.
由于细胞的异质性，从单细胞内源性分子的分析是显着的兴趣，并具有重大意义。虽然显微操作或细胞分选后细胞裂解已经用于随后的分子检测，但直接提取活细胞内容物的方法仍然是实现非破坏性采样和细胞环境保存的具有挑战性但有前途的替代方案。在这里，我们展示了从单细胞的定量提取与时空控制，使用流体力显微镜。我们进一步提出了从细胞质或细胞核中提取的可溶性分子的全面分析，包括酶活性和转录丰度的检测。这种方法揭示了细胞承受高达几皮升提取的能力，并为在单细胞水平上研究生理条件下的细胞动力学和细胞间通讯提供了机会。
Because of cellular heterogeneity, the analysis of endogenous molecules from single cells is of significant interest and has major implications. While micromanipulation or cell sorting followed by cell lysis is already used for subsequent molecular examinations, approaches to directly extract the content of living cells remain a challenging but promising alternative to achieving non-destructive sampling and cell-context preservation. Here, we demonstrate the quantitative extraction from single cells with spatiotemporal control using fluidic force microscopy. We further present a comprehensive analysis of the soluble molecules withdrawn from the cytoplasm or the nucleus, including the detection of enzyme activities and transcript abundances. This approach has uncovered the ability of cells to withstand extraction of up to several picoliters and opens opportunities to study cellular dynamics and cell-cell communication under physiological conditions at the single-cell level.