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.

Next-Generation Microfluidics for Biomedical Research and Healthcare Applications

“…Through 2 Biomedical Engineering and Computational Biology affinity-based multiplexing, the HB-MFP offers insights into cellular phenotypes. In a related field, a non-contact multiphysics probe (NMP) 15 can operate within the physiological tissue environment to precisely manipulate single cells. Through controlled manipulation, electropermeabilization, transfection, and cytoplasm extraction from living single cells, the NMP enhances spatiotemporal single-cell analysis within tissue samples.…”

Less Is More: Oligomer Extraction and Hydrothermal Annealing Increase PDMS Adhesion Forces for Materials Studies and for Biology-Focused Microfluidic Applications

“…Microfluidic devices are a mainstay for the on-chip miniaturization of chemical and biological systems owing to the highly controlled spatiotemporal manipulations of minute sample volumes [ 1 , 2 , 3 ]. Microfluidics are particularly beneficial for resolving mechanisms of the growth, differentiation, and signaling of biological systems that are complex and dynamic [ 4 ].…”

On-Chip Label-Free Sorting of Living and Dead Cells

“…3,4 In addition, in the process of monoclonal antibody production, dead cells can hinder the growth of adjacent living cells and reduce the total production of antibodies. 5 In the emerging field of bio-microfluidics, on-chip sorting of living and dead cells is also a necessary step, as many operations on cells, such as single-cell manipulation, 6,7 cell filtering and cell washing, 8 and removal of cell cryoprotectants, 9 may lead to the presence of some dead cells, which may affect subsequent on-chip experiments.…”

“…For example, in cell-based medical therapies, dead cells have to be removed in advance as they can lead to a series of problems such as host inflammation, which may reduce the therapeutic effect and efficiency of cell therapy. , In drug testing, dead cells also need to be removed as the presence of dead cells reduces the sensitivity of cell-based screening analysis, increases nonspecific binding with reagents, and causes false positive results. , In addition, in the process of monoclonal antibody production, dead cells can hinder the growth of adjacent living cells and reduce the total production of antibodies . In the emerging field of bio-microfluidics, on-chip sorting of living and dead cells is also a necessary step, as many operations on cells, such as single-cell manipulation, , cell filtering and cell washing, and removal of cell cryoprotectants, may lead to the presence of some dead cells, which may affect subsequent on-chip experiments.…”