The Experts below are selected from a list of 20976 Experts worldwide ranked by ideXlab platform
Gwobin Lee - One of the best experts on this subject based on the ideXlab platform.
-
an integrated Microfluidic Platform to perform uninterrupted selex cycles to screen affinity reagents specific to cardiovascular biomarkers
Biosensors and Bioelectronics, 2018Co-Authors: Anirban Sinha, Priya Gopinathan, Yida Chung, Hsinying Lin, Pochiun Huang, Shu Chu Shiesh, Gwobin LeeAbstract:As cardiovascular diseases (CVD) are responsible for millions of deaths annually, there is a need for rapid and sensitive diagnosis of CVD at earlier stages. Aptamers generated by systematic evolution of ligands by exponential enrichment (SELEX) processes have been shown to be superior to conventional antibody-based cardiac biomarker detection. However, SELEX is a complicated, lengthy procedure requiring multiple rounds of extraction/amplification and well-trained personnel. To circumvent such issue, we designed an automated, miniaturized SELEX Platform for the screening of aptamers towards three protein biomarkers associated with CVDs: N-terminal pro-peptide of B-type natriuretic peptide, human cardiac troponin I, and fibrinogen. The developed Microfluidic Platform was equipped with Microfluidic devices capable of sample transport and mixing along with an on-chip nucleic acid amplification module such that the entire screening process (5 rounds of selection in 8 h.) could be performed consecutively on a single chip while consuming only 35 µL of reagents in each cycle. This system may therefore serve as a promising, sensitive, cost-effective Platform for the selection of aptamers specific for CVD biomarkers.
-
high purity and label free isolation of circulating tumor cells ctcs in a Microfluidic Platform by using optically induced dielectrophoretic odep force
Lab on a Chip, 2013Co-Authors: Songbin Huang, Yenheng Lin, Chiahsun Hsieh, Chihliang Yang, Hungchih Lin, Chingping Tseng, Gwobin LeeAbstract:Negative selection-based circulating tumor cell (CTC) isolation is believed valuable to harvest more native, and in particular all possible CTCs without biases relevant to the properties of surface antigens on the CTCs. Under such a cell isolation strategy, however, the CTC purity is normally compromised. To address this issue, this study reports the integration of optically-induced-dielectrophoretic (ODEP) force-based cell manipulation, and a laminar flow regime in a Microfluidic Platform for the isolation of untreated, and highly pure CTCs after conventional negative selection-based CTC isolation. In the design, six sections of moving light-bar screens were continuously and simultaneously exerted in two parallel laminar flows to concurrently separate the cancer cells from the leukocytes based on their size difference and electric properties. The separated cell populations were further partitioned, delivered, and collected through the two flows. With this approach, the cancer cells can be isolated in a continuous, effective, and efficient manner. In this study, the operating conditions of ODEP for the manipulation of prostate cancer (PC-3) and human oral cancer (OEC-M1) cells, and leukocytes with minor cell aggregation phenomenon were first characterized. Moreover, performances of the proposed method for the isolation of cancer cells were experimentally investigated. The results showed that the presented CTC isolation scheme was able to isolate PC-3 cells or OEC-M1 cells from a leukocyte background with high recovery rate (PC-3 cells: 76–83%, OEC-M1 cells: 61–68%), and high purity (PC-3 cells: 74–82%, OEC-M1 cells: 64–66%) (set flow rate: 0.1 μl min−1 and sample volume: 1 μl). The latter is beyond what is currently possible in the conventional CTC isolations. Moreover, the viability of isolated cancer cells was evaluated to be as high as 94 ± 2%, and 95 ± 3% for the PC-3, and OEC-M1 cells, respectively. Furthermore, the isolated cancer cells were also shown to preserve their proliferative capability. As a whole, this study has presented an ODEP-based Microfluidic Platform that is capable of isolating CTCs in a continuous, label-free, cell-friendly, and particularly highly pure manner. All these traits are found particularly meaningful for exploiting the harvested CTCs for the subsequent cell-based, or biochemical assays.
Hadi Shafiee - One of the best experts on this subject based on the ideXlab platform.
-
a Microfluidic Platform for drug screening in a 3d cancer microenvironment
Biosensors and Bioelectronics, 2017Co-Authors: Hardik J Pandya, Karan Dhingra, Devbalaji Prabhakar, Vineethkrishna Chandrasekar, Siva Kumar Natarajan, Anish Vasan, Ashish Kulkarni, Hadi ShafieeAbstract:Development of resistance to chemotherapy treatments is a major challenge in the battle against cancer. Although a vast repertoire of chemotherapeutics is currently available for treating cancer, a technique for rapidly identifying the right drug based on the chemo-resistivity of the cancer cells is not available and it currently takes weeks to months to evaluate the response of cancer patients to a drug. A sensitive, low-cost diagnostic assay capable of rapidly evaluating the effect of a series of drugs on cancer cells can significantly change the paradigm in cancer treatment management. Integration of Microfluidics and electrical sensing modality in a 3D tumour microenvironment may provide a powerful Platform to tackle this issue. Here, we report a 3D Microfluidic Platform that could be potentially used for a real-time deterministic analysis of the success rate of a chemotherapeutic drug in less than 12h. The Platform (66mm×50mm; L×W) is integrated with the microsensors (interdigitated gold electrodes with width and spacing 10µm) that can measure the change in the electrical response of cancer cells seeded in a 3D extra cellular matrix when a chemotherapeutic drug is flown next to the matrix. B16-F10 mouse melanoma, 4T1 mouse breast cancer, and DU 145 human prostate cancer cells were used as clinical models. The change in impedance magnitude on flowing chemotherapeutics drugs measured at 12h for drug-susceptible and drug tolerant breast cancer cells compared to control were 50,552±144 Ω and 28,786±233 Ω, respectively, while that of drug-susceptible melanoma cells were 40,197±222 Ω and 4069±79 Ω, respectively. In case of prostate cancer the impedance change between susceptible and resistant cells were 8971±1515 Ω and 3281±429 Ω, respectively, which demonstrated that the Microfluidic Platform was capable of delineating drug susceptible cells, drug tolerant, and drug resistant cells in less than 12h.
Songbin Huang - One of the best experts on this subject based on the ideXlab platform.
-
label free live and dead cell separation method using a high efficiency optically induced dielectrophoretic odep force based Microfluidic Platform
International Journal of Automation and Smart Technology, 2014Co-Authors: Songbin Huang, Shinglun LiuAbstract:This study reports an optically-induced dielectrophoretic (ODEP) force-based Microfluidic Platform for live and dead cell separation and collection. ODEP forces are used to separate the live and dead cells due to their opposite responses to an ODEP force. Combining the flow control in a Microfluidic system, the live and dead cells can be separated and subsequently collected in an efficient and effective manner. The operating conditions of the ODEP force for manipulating the live and dead chondrocytes is characterized, and separation performance is experimentally evaluated. Results revealed that an applied voltage of 8 V resulted in a maximum difference of manipulation force for the live (49.4 pN) and dead (-20.1 pN) cells. Results of further separation experiments showed that the recovery rate and purity of the isolated live cells was as high as 78.3±6.8 % and 96.4 ±2.2 %, respectively. Overall, the proposed method is found to be particularly valuable for biological research which requires the isolation of highly pure live or dead cells.
-
a clogging free Microfluidic Platform with an incorporated pneumatically driven membrane based active valve enabling specific membrane capacitance and cytoplasm conductivity characterization of single cells
Sensors and Actuators B-chemical, 2014Co-Authors: Songbin Huang, Yang Zhao, Deyong Chen, Tzukeng Chiu, Junbo Wang, Jian Chen, Minhsien WuAbstract:a b s t r a c t This study reports a Microfluidic Platform for clogging-free electrical property analysis of single cells. A pneumatically driven membrane-based active valve was integrated in this Platform to unblock clogging events of constriction microchannels where pneumatic pressures were used to tune the deformation of a polydimethylsiloxan (PDMS) membrane serving as one wall of the constriction microchannel. The proposed Platform was first tested to unblock trapped polystyrene beads at the entrance of constriction microchannels and then the characterization of the cellular electrical properties of lung cancer cells was successfully demonstrated. Results showed that the measured cytoplasm conductivity (0.74 ± 0.20 S/m) and specific membrane capacitance (2.17 ± 0.58 F/cm2) of cells were consistent with the results from the previous publications (0.73 ± 0.17 S/m, and 2.00 ± 0.60 F/cm 2 , respectively). Overall, this study has presented a Microfluidic Platform for single cell analysis with an enhanced function for unblocking cell aggregates at the entrance of microchannels.
-
high purity and label free isolation of circulating tumor cells ctcs in a Microfluidic Platform by using optically induced dielectrophoretic odep force
Lab on a Chip, 2013Co-Authors: Songbin Huang, Yenheng Lin, Chiahsun Hsieh, Chihliang Yang, Hungchih Lin, Chingping Tseng, Gwobin LeeAbstract:Negative selection-based circulating tumor cell (CTC) isolation is believed valuable to harvest more native, and in particular all possible CTCs without biases relevant to the properties of surface antigens on the CTCs. Under such a cell isolation strategy, however, the CTC purity is normally compromised. To address this issue, this study reports the integration of optically-induced-dielectrophoretic (ODEP) force-based cell manipulation, and a laminar flow regime in a Microfluidic Platform for the isolation of untreated, and highly pure CTCs after conventional negative selection-based CTC isolation. In the design, six sections of moving light-bar screens were continuously and simultaneously exerted in two parallel laminar flows to concurrently separate the cancer cells from the leukocytes based on their size difference and electric properties. The separated cell populations were further partitioned, delivered, and collected through the two flows. With this approach, the cancer cells can be isolated in a continuous, effective, and efficient manner. In this study, the operating conditions of ODEP for the manipulation of prostate cancer (PC-3) and human oral cancer (OEC-M1) cells, and leukocytes with minor cell aggregation phenomenon were first characterized. Moreover, performances of the proposed method for the isolation of cancer cells were experimentally investigated. The results showed that the presented CTC isolation scheme was able to isolate PC-3 cells or OEC-M1 cells from a leukocyte background with high recovery rate (PC-3 cells: 76–83%, OEC-M1 cells: 61–68%), and high purity (PC-3 cells: 74–82%, OEC-M1 cells: 64–66%) (set flow rate: 0.1 μl min−1 and sample volume: 1 μl). The latter is beyond what is currently possible in the conventional CTC isolations. Moreover, the viability of isolated cancer cells was evaluated to be as high as 94 ± 2%, and 95 ± 3% for the PC-3, and OEC-M1 cells, respectively. Furthermore, the isolated cancer cells were also shown to preserve their proliferative capability. As a whole, this study has presented an ODEP-based Microfluidic Platform that is capable of isolating CTCs in a continuous, label-free, cell-friendly, and particularly highly pure manner. All these traits are found particularly meaningful for exploiting the harvested CTCs for the subsequent cell-based, or biochemical assays.
Junbo Wang - One of the best experts on this subject based on the ideXlab platform.
-
development of Microfluidic Platform to high throughput quantify single cell intrinsic bioelectrical markers of tumor cell lines subtypes and patient tumor cells
Sensors and Actuators B-chemical, 2020Co-Authors: Yi Zhang, Deyong Chen, Junbo Wang, Hongyan Liang, Huiwen Tan, Yixiang Wang, Jian ChenAbstract:Abstract Although intrinsic bioelectrical markers of single cells (e.g. specific membrane capacitance of Csm, cytoplasm conductivity of σcy and cell diameter of Dc) play key roles in cell-type classification and cell-status evaluation, due to technical limitations, data from large populations of single cells were not available. This paper presents a Microfluidic Platform to high-throughput quantify single-cell intrinsic bioelectrical markers of tumor cell lines, subtypes and patient tumor cells, where an asymmetrical constriction channel defined by a major and two side constriction channels was used as the sensitive unit. The developed Microfluidic Platform was used to quantify ∼100,000 single-cell Csm, σcy and Dc from (I) eight tumor cell lines of A549, Hep G2, SW620, AGS, PANC-1, Hela, CAL 27 and HL-60 with cell-type classification rates of 87.6 %±9.1 % obtained; (II) tumor cell subtypes of 293 T with and without the transfection of the CRISPR knockout library where different distributions of Csm were located; (III) patient tumor cells with classification rates of 96.1 %±3.8 % obtained compared with tumor cell lines. In conclusion, the Microfluidic Platform developed in this study can function as a high-throughput tool in the field of single-cell analysis.
-
development of Microfluidic Platform capable of high throughput absolute quantification of single cell multiple intracellular proteins from tumor cell lines and patient tumor samples
Biosensors and Bioelectronics, 2020Co-Authors: Lixing Liu, Deyong Chen, Yixiang Wang, Hongyu Yang, Dong Men, Meng Wang, Xiaolei Gao, Ting Zhang, Chunlai Xue, Junbo WangAbstract:Quantification of single-cell proteins plays key roles in cell heterogeneity while due to technical limitations absolute numbers of multiple intracellular proteins from large populations of single cells were still missing, leading to compromised results in cell-type classifications. This paper presents a Microfluidic Platform capable of high-throughput absolute quantification of single-cell multiple types of intracellular proteins where cells stained with fluorescent labelled antibodies are aspirated into the constriction microchannels with excited fluorescent signals detected and translated into numbers of binding sites of targeted proteins based on calibration curves formed by flushing gradient solutions of fluorescent labelled antibodies directly into constriction microchannels. Based on this approach, single-cell numbers of binding sites of β-actin, α-tubulin and β-tubulin from tens of thousands of five representative tumor cell lines were first quantified, reporting cell-type classification rates of 83.0 ± 7.1%. Then single-cell numbers of binding sites of β-actin, biotin and RhoA from thousands of five tumor cell lines with varieties in malignant levels were quantified, reporting cell-type classification rates of 93.7 ± 2.8%. Furthermore, single-cell numbers of binding sites of Ras, c-Myc and p53 from thousands of cells derived from two oral tumor lines of CAL 27, WSU-HN6 and two oral tumor patient samples were quantified, contributing to high classifications of both tumor cell lines (98.6%) and tumor patient samples (83.4%). In conclusion, the developed Microfluidic Platform was capable of quantifying multiple intracellular proteins from large populations of single cells, and the collected data of protein expressions enabled effective cell-type classifications.
-
a clogging free Microfluidic Platform with an incorporated pneumatically driven membrane based active valve enabling specific membrane capacitance and cytoplasm conductivity characterization of single cells
Sensors and Actuators B-chemical, 2014Co-Authors: Songbin Huang, Yang Zhao, Deyong Chen, Tzukeng Chiu, Junbo Wang, Jian Chen, Minhsien WuAbstract:a b s t r a c t This study reports a Microfluidic Platform for clogging-free electrical property analysis of single cells. A pneumatically driven membrane-based active valve was integrated in this Platform to unblock clogging events of constriction microchannels where pneumatic pressures were used to tune the deformation of a polydimethylsiloxan (PDMS) membrane serving as one wall of the constriction microchannel. The proposed Platform was first tested to unblock trapped polystyrene beads at the entrance of constriction microchannels and then the characterization of the cellular electrical properties of lung cancer cells was successfully demonstrated. Results showed that the measured cytoplasm conductivity (0.74 ± 0.20 S/m) and specific membrane capacitance (2.17 ± 0.58 F/cm2) of cells were consistent with the results from the previous publications (0.73 ± 0.17 S/m, and 2.00 ± 0.60 F/cm 2 , respectively). Overall, this study has presented a Microfluidic Platform for single cell analysis with an enhanced function for unblocking cell aggregates at the entrance of microchannels.
Sebastian J Maerkl - One of the best experts on this subject based on the ideXlab platform.
-
a digital analog Microfluidic Platform for patient centric multiplexed biomarker diagnostics of ultralow volume samples
ACS Nano, 2016Co-Authors: Francesco Piraino, Francesca Volpetti, Craig Watson, Sebastian J MaerklAbstract:Microfluidic diagnostic devices have the potential to transform the practice of medicine. We engineered a multiplexed digital–analog Microfluidic Platform for the rapid and highly sensitive detection of 3–4 biomarkers in quadruplicate in 16 independent and isolated Microfluidic unit cells requiring only a single 5 μL sample. We comprehensively characterized the Platform by performing single enzyme and digital immunoassays, achieving single molecule detection and measured as low as ∼10 fM (330 fg/mL) GFP in buffer and ∼12 fM GFP in human serum. We applied our integrated digital detection mechanism to multiplexed detection of 1pM anti-Ebola IgG in human serum and were able to differentiate three common Ebola strains. To ascertain that the device can be applied in environments beyond clinical point-of-care settings, we developed a low-cost, portable hardware system to control and read out the Microfluidic device and detected anti-Ebola IgG in ultralow volume whole blood samples to levels of 100 pM in a multi...
-
a digital analog Microfluidic Platform for patient centric multiplexed biomarker diagnostics of ultralow volume samples
ACS Nano, 2016Co-Authors: Francesco Piraino, Francesca Volpetti, Craig Watson, Sebastian J MaerklAbstract:Microfluidic diagnostic devices have the potential to transform the practice of medicine. We engineered a multiplexed digital-analog Microfluidic Platform for the rapid and highly sensitive detection of 3-4 biomarkers in quadruplicate in 16 independent and isolated Microfluidic unit cells requiring only a single 5 μL sample. We comprehensively characterized the Platform by performing single enzyme and digital immunoassays, achieving single molecule detection and measured as low as ∼10 fM (330 fg/mL) GFP in buffer and ∼12 fM GFP in human serum. We applied our integrated digital detection mechanism to multiplexed detection of 1pM anti-Ebola IgG in human serum and were able to differentiate three common Ebola strains. To ascertain that the device can be applied in environments beyond clinical point-of-care settings, we developed a low-cost, portable hardware system to control and read out the Microfluidic device and detected anti-Ebola IgG in ultralow volume whole blood samples to levels of 100 pM in a multiplexed assay format.
