Noncontact Multiphysics Probe for Spatiotemporal Resolved Single‐Cell Manipulation and Analyses

Abstract 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.