Deyong Chen

Microfluidic impedance flow cytometry enabling high-throughput single-cell electrical property characterization

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.

A microfluidic system enabling continuous characterization of specific membrane capacitance and cytoplasm conductivity of single cells in suspension

This paper presents a microfluidic system enabling continuous characterization of specific membrane capacitance (Cspecific membrane) and cytoplasm conductivity (σcytoplasm) of single cells in suspension. In this study, cells were aspirated continuously through a constriction channel while cell elongations and impedance profiles at two frequencies (1kHz and 100kHz) were measured simultaneously using microscopy imaging and a lock-in amplifier. 1kHz impedance data were used to evaluate cellular sealing properties with constriction channel walls and 100kHz impedance data were translated to quantify equivalent membrane capacitance and cytoplasm resistance of single cells, which were further translated to Cspecific membrane and σcytoplasm. Two model cell lines (kidney tumor cell line of 786-O (n=302) and vascular smooth muscle cell line of T2 (n=216)) were used to evaluate this technique, producing Cspecific membrane of 3.67±1.00 and 4.53±1.51μF/cm(2) and σcytoplasm of 0.47±0.09 and 0.55±0.14S/m, respectively. Compared to previously reported techniques which can only collect Cspecific membrane and σcytoplasm from tens of cells, this new technique has a higher throughput, capable of collecting Cspecific membrane and σcytoplasm from hundreds of cells in 30min immediately after cell passage.

A MEMS Based Electrochemical Vibration Sensor for Seismic Motion Monitoring

This paper presents a micro-electro-mechanical system (MEMS) based electrochemical vibration sensor for seismic detection. Simulations were conducted to analyze the effect of the insulating spacers thickness on the sensitivity of the devices. Then, devices with different insulating spacers thicknesses were fabricated based on MEMS processes. The devices sensitivity was confirmed by a customized experimental platform, verifying the simulation results. In addition, the device performance was characterized with a quantified bandwidth of 0.2-5 Hz with sensitivity of 30.2 V/(m/s2) and a linear voltage output as a function of the input vibration amplitude (up to 10 mg). The devices noise was determined as -140 dB at 1 Hz. A random-vibration testing in the laboratory environment was conducted, where response correlations among seven devices were calculated as 0.976±0.017, suggesting high device repeatability. A field test was conducted right above a subway line to detect the seismic motion caused by the train and test results showed that the performance of the proposed devices was comparable with that of the commercial product MET-1001. A field test was also conducted in a prairie to monitor natural seismic motions and experimental results indicated that the proposed electrochemical sensors can detect seismic motions in a lower frequency domain with an energy peak at around 0.3 Hz compared with conventional moving-coil seismic sensors with an energy peak at 3 Hz. This newly proposed vibration sensor may function as a promising seismic motion detecting device in the field of geophysical prospecting where low-frequency seismic motion detection is requested.

A microfluidic system for cell type classification based on cellular size-independent electrical properties

This paper presents a microfluidic system enabling cell type classification based on continuous characterization of size-independent electrical properties (e.g., specific membrane capacitance (C(specific membrane)) and cytoplasm conductivity (σ(cytoplasm)). In this study, cells were aspirated continuously through a constriction channel, while cell elongation and impedance profiles at two frequencies (1 kHz and 100 kHz) were measured simultaneously. Based on a proposed distributed equivalent circuit model, 1 kHz impedance data were used to evaluate cellular sealing properties with constriction channel walls and 100 kHz impedance data were translated to C(specific membrane) and σ(cytoplasm). Two lung cancer cell lines of CRL-5803 cells (n(cell) = 489) and CCL-185 cells (n(cell) = 487) were used to evaluate this technique, producing a C(specific membrane) of 1.63 ± 0.52 μF cm(-2) vs. 2.00 ± 0.60 μF cm(-2), and σ(cytoplasm) of 0.90 ± 0.19 S m(-1)vs. 0.73 ± 0.17 S m(-1). Neural network-based pattern recognition was used to classify CRL-5803 and CCL-185 cells, producing success rates of 65.4% (C(specific membrane)), 71.4% (σ(cytoplasm)), and 74.4% (C(specific membrane) and σ(cytoplasm)), suggesting that these two tumor cell lines can be classified based on their electrical properties.

Development of Droplet Microfluidics Enabling High-Throughput Single-Cell Analysis

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.

A High-Q Resonant Pressure Microsensor with Through-Glass Electrical Interconnections Based on Wafer-Level MEMS Vacuum Packaging

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.

Low Frequency Electrochemical Accelerometer with Low Noise Based on MEMS

Petroleum prospecting and early warning of some geological disaster increasingly depend on the accelerometers which can detect vibrate of frequency below 1Hz, but it’s embarrassing that accelerometers based on Si or SiO2 structure make an awful performance in this frequency range. Electrochemical accelerometers were developed in 1990s. With fluidics to be inertial mass, electrochemical accelerometer not only show an excellent property in low frequency, but also has a wide dynamic range. However, traditional fabrication process of electrochemical accelerometer is rather complex and can’t eliminate the noise due to the inconsistency and asymmetry of electrodes. To solve these problems, a scheme based on MEMS is proposed here, including design, fabrication and package. Properties of electrochemical accelerometer (EAM) are tested in two conditions at last.

An Electromagnetically Excited Silicon Nitride Beam Resonant Accelerometer

A resonant microbeam accelerometer of a novel highly symmetric structure based on MEMS bulk-silicon technology is proposed and some numerical modeling results for this scheme are presented. The accelerometer consists of two proof masses, four supporting hinges, two anchors, and a vibrating triple beam, which is clamped at both ends to the two proof masses. LPCVD silicon rich nitride is chosen as the resonant triple beam material, and parameter optimization of the triple-beam structure has been performed. The triple beam is excited and sensed electromagnetically by film electrodes located on the upper surface of the beam. Both simulation and experimental results show that the novel structure increases the scale factor of the resonant accelerometer, and ameliorates other performance issues such as cross axis sensitivity of insensitive input acceleration, etc.

Development of Microfluidic Systems Enabling High-Throughput Single-Cell Protein Characterization

This article reviews recent developments in microfluidic systems enabling high-throughput characterization of single-cell proteins. Four key perspectives of microfluidic platforms are included in this review: (1) microfluidic fluorescent flow cytometry; (2) droplet based microfluidic flow cytometry; (3) large-array micro wells (microengraving); and (4) large-array micro chambers (barcode microchips). We examine the advantages and limitations of each technique and discuss future research opportunities by focusing on three key performance parameters (absolute quantification, sensitivity, and throughput).

Single-Cell Electrical Phenotyping Enabling the Classification of Mouse Tumor Samples

Single-cell electrical phenotyping (e.g., specific membrane capacitance (Cm) and cytoplasm conductivity (σp)) has long been regarded as potential label-free biophysical markers in tumor status evaluation. However, previous studies only reported the differentiation of tumor cell lines without classifying real tumor samples using cellular electrical properties. In this study, two types of mouse tumor models were constructed by injecting two types of tumor cell lines (A549 and H1299), respectively. Then tumor portions were retrieved for immunohistochemistry studies and single-cell electrical phenotyping based on home-developed microfluidic platforms. Immunohistochemistry results of tumor samples confirmed the adenocarcinoma and large-cell carcinoma characteristics for A549 and H1299 based tumor samples, respectively. Meanwhile, cellular Cm and σp were characterized as 2.25 ± 0.50 μF/cm2 and 0.96 ± 0.20 S/m for A549 based tumor samples (ncell = 1336, Mouse I, II, III) and 1.76 ± 0.54 μF/cm2 and 1.35 ± 0.28 S/m for H1299 based tumor samples (ncell = 1442, Mouse IV, V, VI). Significant differences in Cm and σp were observed between these two types of tumor samples, validating the feasibility of using Cm and σp for mouse tumor classification.