This article reviews recent developments in microfluidic impedance flow cytometry for high-throughput electrical property characterization of single cells. Four major perspectives of microfluidic impedance flow cytometry for single-cell characterization are included in this review: (1) early developments of microfluidic impedance flow cytometry for single-cell electrical property characterization; (2) microfluidic impedance flow cytometry with enhanced sensitivity; (3) microfluidic impedance and optical flow cytometry for single-cell analysis and (4) integrated point of care system based on microfluidic impedance flow cytometry. We examine the advantages and limitations of each technique and discuss future research opportunities from the perspectives of both technical innovation and clinical applications.

This article reviews recent developments in droplet microfluidics enabling high-throughput single-cell analysis. Five key aspects in this field are included in this review: (1) prototype demonstration of single-cell encapsulation in microfluidic droplets; (2) technical improvements of single-cell encapsulation in microfluidic droplets; (3) microfluidic droplets enabling single-cell proteomic analysis; (4) microfluidic droplets enabling single-cell genomic analysis; and (5) integrated microfluidic droplet systems enabling single-cell screening. We examine the advantages and limitations of each technique and discuss future research opportunities by focusing on key performances of throughput, multifunctionality, and absolute quantification.

In this paper, a piezoresistive pressure sensor based on silicon on insulator (SOI) was presented, which was composed of an SOI layer with sensing elements and a glass cap for a hermetic package. Different from its conventional counterparts, the position and thickness of the four piezoresistors was optimized based on numerical simulation, which suggests that two piezoresistors at the center while the other two at the edge of the pressure-sensitive diaphragm and a thickness of 2 μm can produce the maximum sensitivity and the minimum nonlinearity. Due to the use of silicon rather than metal for electrical connections, the piezoresistive pressure sensor was fabricated in a highly simplified process. From the experimental results, the fabricated piezoresistive pressure sensor demonstrated a high sensitivity of 37.79 mV·V⁻¹·MPa⁻¹, a high full-scale (FS) output of 472.33 mV, a low hysteresis of 0.09% FS, a good repeatability of 0.03% FS and a good accuracy of 0.06% FS at 20 °C. A temperature coefficient of sensitivity of 0.44 mV·MPa⁻¹·°C⁻¹ and a low zero drift were also shown at different temperatures. The piezoresistive pressure sensor developed in this study may function as an enabling tool in pressure measurements.

This paper presents a high-Q resonant pressure microsensor with through-glass electrical interconnections based on wafer-level MEMS vacuum packaging. An approach to maintaining high-vacuum conditions by integrating the MEMS fabrication process with getter material preparation is presented in this paper. In this device, the pressure under measurement causes a deflection of a pressure-sensitive silicon square diaphragm, which is further translated to stress build up in “H” type doubly-clamped micro resonant beams, leading to a resonance frequency shift. The device geometries were optimized using FEM simulation and a 4-inch SOI wafer was used for device fabrication, which required only three photolithographic steps. In the device fabrication, a non-evaporable metal thin film as the getter material was sputtered on a Pyrex 7740 glass wafer, which was then anodically bonded to the patterned SOI wafer for vacuum packaging. Through-glass via holes predefined in the glass wafer functioned as the electrical interconnections between the patterned SOI wafer and the surrounding electrical components. Experimental results recorded that the Q-factor of the resonant beam was beyond 22,000, with a differential sensitivity of 89.86 Hz/kPa, a device resolution of 10 Pa and a nonlinearity of 0.02% F.S with the pressure varying from 50 kPa to 100 kPa. In addition, the temperature drift coefficient was less than −0.01% F.S/°C in the range of −40 °C to 70 °C, the long-term stability error was quantified as 0.01% F.S over a 5-month period and the accuracy of the microsensor was better than 0.01% F.S.

This letter presents a MEMS resonant differential pressure sensor with an integrated temperature sensor and a static pressure sensor to address the issue of side effects of temperature and static pressure on measurement accuracies of resonant differential pressure microsensors. In this microsensor, two resonators were employed on the pressure sensitive diaphragm to sense differential pressure while a Pt-film-resistor was included as the temperature sensor and an additional resonator was positioned on the framework as the static pressure sensor. Microfabrication was conducted to manufacture the resonant microsensor and key performances were quantified as 79.76 Hz/kPa (~1036 ppm/kPa) of differential pressure sensitivity, 6.22 Ω/°C of temperature sensor sensitivity and –0.47 Hz/kPa of static pressure sensor sensitivity. Based on compensations of temperature and static pressure, a measurement accuracy of 0.02%FS was achieved for the developed microsensor under the differential pressure range from 0 to 100 kPa, the temperature range from –20°C to 60°C and the static pressure range from 110 kPa to 200 kPa.

Quantification of single-cell proteomics provides key insights into cellular heterogeneity while conventional flow cytometry cannot provide absolute quantification of intracellular proteins of single cells due to the lack of calibration approaches. This paper presents a constriction channel (with a cross sectional area smaller than cells) based microfluidic flow cytometer, capable of collecting copy numbers of specific intracellular proteins. In this platform, single cells stained with fluorescence labelled antibodies were forced to squeeze through the constriction channel with the fluorescence intensities quantified and since cells fully filled the constriction channel during the squeezing process, solutions with fluorescence labelled antibodies were flushed into the constriction channel to obtain calibration curves. By combining raw fluorescence data and calibration curves, absolute quantification of intracellular proteins was realized. As a demonstration, copy numbers of beta-actin of single tumour cells were quantified to be 0.90 ± 0.30 μM (A549, n_cell = 14 228), 2.34 ± 0.70 μM (MCF 10A, n_cell = 2455), and 0.98 ± 0.65 μM (Hep G2, n_cell = 6945). The travelling time for individual cells was quantified to be roughly 10 ms and thus a throughput of 100 cells per s can be achieved. This microfluidic system can be used to quantify the copy numbers of intracellular proteins in a high-throughput manner, which may function as an enabling technique in the field of single-cell proteomics.