Noncontact Multiphysics Probe for Spatiotemporal Resolved Single-Cell Manipulation and Analyses
Noncontact Multiphysics Probe for Spatiotemporal Resolved Single-Cell Manipulation and Analyses
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DOI：
10.1002/smll.202100801
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发表时间：
2021-05-19
期刊：
影响因子：
13.3
通讯作者：
Qasaimeh, Mohammad A.
中科院分区：
文献类型：
--
作者：
Brimmo, Ayoola T.;Menachery, Anoop;Qasaimeh, Mohammad A.
组织内单个细胞的异质性和空间排列对于宿主多细胞生物体的身份至关重要。虽然目前的单细胞技术能够解决异质性，但它们主要依赖于从其生理环境中提取靶细胞，因此失去了理解细胞网络所需的时空分辨率。在这里，一个多功能的非接触式扫描探针，可以精确地执行多个操作程序的活单细胞，而在其生理组织环境中，被证明。非接触式多物理场探针（NMP）由流体孔和“驼峰”形电极组成，它们以单细胞分辨率同时限制试剂和电信号。NMP的独特的电渗透为基础的方法，通过细胞膜转移大分子。通过用不同的DNA质粒载体分离相邻的单细胞，证明了该技术的可调节空间能力。NMP技术还为从活单细胞中可控地提取细胞质打开了大门。通过对粘附细胞进行多个时间点活检而不影响提取的大分子的完整性或细胞的活力来证明这种强大的应用。此外，NMP的功能，作为一个电热为基础的微流控全细胞镊子的报告。这项工作提供了一个多功能的工具，具有前所未有的探测功能，用于组织样本内的时空单细胞分析。
Heterogeneity and spatial arrangement of individual cells within tissues are critical to the identity of the host multicellular organism. While current single-cell techniques are capable of resolving heterogeneity, they mostly rely on extracting target cells from their physiological environment and hence lose the spatiotemporal resolution required for understanding cellular networks. Here, a multifunctional noncontact scanning probe that can precisely perform multiple manipulation procedures on living single-cells, while within their physiological tissue environment, is demonstrated. The noncontact multiphysics probe (NMP) consists of fluidic apertures and "hump" shaped electrodes that simultaneously confine reagents and electric signals with a single-cell resolution. The NMP's unique electropermealization-based approach in transferring macromolecules through the cell membrane is presented. The technology's adjustable spatial ability is demonstrated by transfecting adjacent single-cells with different DNA plasmid vectors. The NMP technology also opens the door for controllable cytoplasm extraction from living single-cells. This powerful application is demonstrated by executing multiple time point biopsies on adherent cells without affecting the integrity of the extracted macromolecules or the viability of cells. Furthermore, the NMP's function as an electro-thermal based microfluidic whole-cell tweezer is reported. This work offers a multifunctional tool with unprecedented probing features for spatiotemporal single-cell analysis within tissue samples.