3D rotation is one of many fundamental manipulations to cells and imperative in a wide range of applications in single cell analysis involving biology, chemistry, physics and medicine. In this article, we report a dielectrophoresis-based, on-chip manipulation method that can load and rotate a single cell for 3D cell imaging and multiple biophysical property measurements. To achieve this, we trapped a single cell in constriction and subsequently released it to a rotation chamber formed by four sidewall electrodes and one transparent bottom electrode. In the rotation chamber, rotating electric fields were generated by applying appropriate AC signals to the electrodes for driving the single cell to rotate in 3D under control. The rotation spectrum for in-plane rotation was used to extract the cellular dielectric properties based on a spherical single-shell model, and the stacked images of out-of-plane cell rotation were used to reconstruct the 3D cell morphology to determine its geometric parameters. We have tested the capabilities of our method by rotating four representative mammalian cells including HeLa, C3H10, B lymphocyte, and HepaRG. Using our device, we quantified the area-specific membrane capacitance and cytoplasm conductivity for the four cells, and revealed the subtle difference of geometric parameters (i.e., surface area, volume, and roughness) by 3D cell imaging of cancer cells and normal leukocytes. Combining microfluidics, dielectrophoresis, and microscopic imaging techniques, our electrorotation-on-chip (EOC) technique is a versatile method for manipulating single cells under investigation and measuring their multiple biophysical properties.

Single-cell impedance measurement is a label-free, noninvasive method for characterizing the electrical properties of single cells. At present, though widely used for impedance measurement, electric impedance flow cytometry (IFC) and electric impedance spectroscopy (EIS) are used alone for most microfluidic chips. In this paper, we present a microfluidic device combining the IFC and EIS techniques for single-cell electrical property measurement. The device uses hydrodynamic constriction to passively trap single cells and uses coplanar electrodes to obtain the impedance spectrum of the trapped cell via EIS and discrete impedance data points of the passing cells via IFC. Through experiment, we verified the individual functionality of IFC and EIS respectively, by revealing through IFC the impedance magnitude difference and quantifying through EIS the area-specific membrane capacitance and cytoplasm conductivity of the three types of cancer cells. We also demonstrated the complementarity of IFC and EIS, which holds for a wide range of the flow rate. We envision that the strategy of combining IFC and EIS provides a new thought in the efforts to enhancing the efficiency of electrical property measurement for single cells.

Cells have different intrinsic markers such as mechanical and electrical properties, which may be used as specific characteristics. Here, we present a microfluidic chip configured with two opposing optical fibers and four 3D electrodes for multiphysical parameter measurement. The chip leverages optical fibers to capture and stretch a single cell and uses 3D electrodes to achieve rotation of the single cell. According to the stretching deformation and rotation spectrum, the mechanical and dielectric properties can be extracted. We provided proof of concept by testing five types of cells (HeLa, A549, HepaRG, MCF7 and MCF10A) and determined five biophysical parameters, namely, shear modulus, steady-state viscosity, and relaxation time from the stretching deformation and area-specific membrane capacitance and cytoplasm conductivity from the rotation spectra. We showed the potential of the chip in cancer research by observing subtle changes in the cellular properties of transforming growth factor beta 1 (TGF-β1)-induced epithelial–mesenchymal transition (EMT) A549 cells. The new chip provides a microfluidic platform capable of multiparameter characterization of single cells, which can play an important role in the field of single-cell research.

Single cell analysis has received increasing attention recently in both academia and clinics, and there is an urgent need for effective upstream cell sample preparation. Two extremely challenging tasks in cell sample preparation-highefficiency cell enrichment and precise single cell capture-have now entered into an era full of exciting technological advances, which are mostly enabled by microfluidics. In this review, we summarize the category of technologies that provide new solutions and creative insights into the two tasks of cell manipulation, with a focus on the latest development in the recent five years by highlighting the representative works. By doing so, we aim both to outline the framework and to showcase example applications of each task. In most cases for cell enrichment, we take circulating tumor cells (CTCs) as the target cells because of their research and clinical importance in cancer. For single cell capture, we review related technologies for many kinds of target cells because the technologies are supposed to be more universal to all cells rather than CTCs. Most of the mentioned technologies can be used for both cell enrichment and precise single cell capture. Each technology has its own advantages and specific challenges, which provide opportunities for researchers in their own area. Overall, these technologies have shown great promise and now evolve into real clinical applications. Published by AIP Publishing. [http://dx

In this study, we proposed a microfluidic device with compact structures integrating multiple modalities for cell capture, pairing, fusion, and culture. The microfluidic device is composed of upper and lower parts. The lower part configured with electrodes and capture wells is used for cell trapping/pairing/fusion, while the upper part configured with corresponding culture wells is used for cell culture. Dielectrophoresis is used to enable accurate cell trapping and pairing in capture wells. Moreover, the paired cells are fused flexibly by either electrical pulses or polyethylene glycol (PEG) buffer. The fused cells are then transferred to culture wells for on-chip culture simply by flipping the device. Using the device and HeLa cells, we demonstrated pairing efficiency of ∼78% and fusion efficiencies of ∼26% for electrical fusion or ∼21% for PEG fusion, and successful cell proliferation and migration after 72 h on-chip culture. We believe that this multifunction-integrated but structure-simplified microfluidic device would largely facilitate cell fusion oriented tasks.