Ce Wang's research while affiliated with Suzhou Institute of Biomedical Engineering and Technology, Chinese Academy of Sciences and other places

Small extracellular vesicles (sEVs), a form of extracellular vesicles, are lipid bilayered structures released by all cells. Large-scale studies on sEVs from clinical samples are necessary, but a major obstacle is the lack of rapid, reproducible, efficient, and low-cost methods to enrich sEVs. Acoustic microfluidics have the advantage of being label-free and biocompatible, which have been reported to successfully enrich sEVs. In this paper, we present a highly efficient acoustic microfluidic trap that can offer low and large volume compatible ways of enriching sEVs from biological fluids by flexible structure design. It uses the idea of pre-loading larger seed particles in the acoustic trap to enable sub-micron particle capturing. The microfluidic chip is actuated using a piezoelectric plate transducer attached to a silicon-glass bonding plate with circular cavities. Each cavity works as a resonant unit, excited at the frequency of both the half wave resonance in the main plane and inverted quarter wave resonance in the depth direction, which has the ability to strongly trap seed particles at the center, thereby improving the subsequent nanoparticle capture efficiency. Mean trapping efficiencies of 35.62% and 64.27% were obtained using 60 nm and 100 nm nanobeads, respectively. By the use of this technology, we have successfully enriched sEVs from cell culture conditioned media and blood plasma at a flow rate of 10 μL min-1. The isolated sEV subpopulations are characterized by NTA and TEM, and their protein cargo is determined by WB. This acoustic trapping chip provides a rapid and robust method to enrich sEVs from biofluids with high reproducibility and sufficient quantities. Therefore, it can serve as a new tool for biological and clinical research such as cancer diagnosis and drug delivery.

Raman flow cytometry (RFC) uniquely integrates the “label-free” capability of Raman spectroscopy with the “high-throughput” attribute of traditional flow cytometry (FCM), offering exceptional performance in cell characterization and sorting. Unlike conventional FCM, RFC stands out for its elimination of the dependency on fluorescent labels, thereby reducing interference with the natural state of cells. Furthermore, it significantly enhances the detection information, providing a more comprehensive chemical fingerprint of cells. This review thoroughly discusses the fundamental principles and technological advantages of RFC and elaborates on its various applications in the biomedical field, from identifying and characterizing cancer cells for in vivo cancer detection and surveillance to sorting stem cells, paving the way for cell therapy, and identifying metabolic products of microbial cells, enabling the differentiation of microbial subgroups. Moreover, we delve into the current challenges and future directions regarding the improvement in sensitivity and throughput. This holds significant implications for the field of cell analysis, especially for the advancement of metabolomics.

Skin wounds, especially large-area skin trauma, would bring great pain and even fatal risk to patients. In recent years, local autologous cell transplantation has shown great potential for wound healing and re-epithelialization. However, when the cell suspension prepared with normal saline is delivered to the wound, due to its low viscosity, it is easy to form big drops in the deposition and lose them from the wound bed, resulting in cell loss and uneven coverage. Here, we developed a novel air-assisted atomization device (AAAD). Under proper atomization parameters, 1% (w/v) sodium alginate (SA) solution carrier could be sprayed uniformly. Compared with normal saline, the run-off of the SA on the surface of porcine skin was greatly reduced. In theory, the spray height of AAAD could be set to achieve the adjustment of a large spray area of 1–12 cm ² . In the measurement of droplet velocity and HaCaT cell viability, the spray height of AAAD would affect the droplet settling velocity and then the cell delivery survival rate (CSR). Compared with the spray height of 50 mm, the CSR of 100 mm was significantly higher and could reach 91.09% ± 1.82% (92.82% ± 2.15% in control). For bio-ink prepared with 1% (w/v) SA, the viability remained the same during the 72-h incubation. Overall, the novel AAAD uniformly atomized bio-ink with high viscosity and maintained the viability and proliferation rate during the delivery of living cells. Therefore, AAAD has great potential in cell transplantation therapy, especially for large-area or irregular skin wounds.

Cell enrichment is a powerful tool in many kinds of cell research, especially in applications with low abundance cell types. In this work, we developed a microfluidic fluorescence activated cell sorting (μFACS) device that is able to perform on-demand, low loss cell detection and sorting. The chip utilizes three-dimensional acoustic standing waves to position all cells in the same fluid velocity regime without sheath. When the cells pass through a laser interrogation region, the scattering and fluorescent signals are detected, translated and transported to software. The target cells are then identified by gating on the plots. Short bursts of standing acoustic waves are triggered by order from PC to sort target cells within predefined gating region. For very low abundance and rare labeled lymphocyte mixed with high concentration unlabeled white blood cells (WBCs), (1–100 labeled lymphocytes are diluted in 10⁶ WBCs in 1 mL volume fluid), the device is able to remove more than 98% WBCs and recover labeled lymphocyte with efficiency of 80%. We further demonstrated that this device worked with real clinical samples by successfully isolating fetal nucleated red blood cells (FNRBCs) in the blood samples from pregnant women with male fetus. The obtained cells were sequenced and the expressions of (sex determining region Y) SRY genes were tested to determine fetal cell proportion. In genetic analysis, the proportion of fetal cells in the final picked sample is up to 40.64%. With this ability, the device proposed could be valuable for biomedical applications involving fetal cells, circulating tumor cells, and stem cells. This article is protected by copyright. All rights reserved.

In recent years, microflow cytometry has become a popular research field because of its potential to provide low‐cost and disposable chips for complex cell analyses. Herein, we demonstrate a sheathless microflow cytometer which integrates a bulk standing acoustic wave based microchip capable of three dimensional cell focusing. Flow cytometry was successfully demonstrated using this system with a coefficient of variation (CV) of 2.16% with standard calibration beads. The sensitivities calibrated by rainbow beads are 518 MEFL in FITC channel and 264 MEPE in PE channels respectively. The linearities are more than 99% in both channels. The capability of the proposed microflow cytometer is further demonstrated by immunologically labeled leukocytes differentiation in blood. This acoustic‐based microflow cytometer did not require any sheath flows or complex structures and can be mass produced. Because of the simple fluid channel, the chip can be easily made pipeless, disposable for applications requiring no cross contamination. Moreover, with the gentle and bio‐compatible acoustic waves used, this technique is expected to maintain the viability of cells and other bioparticles. This article is protected by copyright. All rights reserved.

A compact driver based on current feedback amplifiers is designed to drive interdigital transducers (IDTs) that generate standing surface acoustic waves for cell sorting. Compared with commercial RF amplifiers, this driver can be used to drive a wider range of loads without impedance matching. Furthermore, the driver works in a switch mode triggered by target cells, which significantly reduces power consumption in the system. A Butterworth–Van Dyke equivalent circuit was fabricated to study the electrical characteristics of the IDTs, and the driver was designed and optimized by circuit simulations. A cell sorter was constructed and tested experimentally to demonstrate that the driver meets sorting requirements. The driver allows the cell sorter to extract rare cells while otherwise consuming low power.