2013 13th IEEE International Conference on Nanotechnology (IEEE-NANO 2013), 2013
ABSTRACT This paper presents a Nafion film based micro-nanofluidic device for concurrent DNA prec... more ABSTRACT This paper presents a Nafion film based micro-nanofluidic device for concurrent DNA preconcentration and separation. The principle of the device is based on: (a) ion concentration polarization phenomenon at the junction of the microchannel and the nanochannels in the Nafion film to form opposing electrophoretic and electroosmotic forces acting on the DNAs, combined with (b) end-labeled free solution electrophoresis to vary the charge-to-mass ratio for molecular differentiation. Extensive experiments were carried out to characterize the functionality of the device. Concurrent preconcentration and separation of a DNA mixture within 240s were successfully demonstrated, yielding concentration ratios up to 1,150X and separation resolution of 1.85. The effect of applied electric field on the concentration and separation performance was investigated. The device can be used as a key sample preparation element in conjunction with micro- or nanofluidic sensors to obtained microTAS functionality.
Due to the significance of the leukocyte-endothelium interactions in pathogenesis of disease and ... more Due to the significance of the leukocyte-endothelium interactions in pathogenesis of disease and drug delivery, amongst many others, several in vitro models have been developed to study different aspects of the leukocyte adhesion cascade. Current in vitro models of the leukocyte adhesion cascade cannot be used for real-time studies of the entire leukocyte adhesion cascade including rolling, adhesion and migration in a single assay. In this study, we have developed and validated a novel microfluidic assay and used it to test the hypothesis that blocking of specific steps in the adhesion/migration cascade significantly affects other steps of the cascade.
Tumor drug delivery is a complex phenomenon affected by several elements in addition to drug or d... more Tumor drug delivery is a complex phenomenon affected by several elements in addition to drug or delivery vehicle's physico-chemical properties. A key factor is tumor microvasculature with complex effects including convective transport, high interstitial pressure and enhanced vascular permeability due to the presence of "leaky vessels". Current in vitro models of the tumor microenvironment for evaluating drug delivery are oversimplified and, as a result, show poor correlation with in vivo performance. In this study, we report on the development of a novel microfluidic platform that models the tumor microenvironment more accurately, with physiologically and morphologically realistic microvasculature including endothelial cell lined leaky capillary vessels along with 3D solid tumors. Endothelial cells and 3D spheroids of cervical tumor cells were co-cultured in the networks. Drug vehicle screening was demonstrated using GFP gene delivery by different formulations of nanopolymers. The synthetic tumor network was successful in predicting in vivo delivery efficiencies of the drug vehicles. The developed assay will have critical applications both in basic research, where it can be used to develop next generation delivery vehicles, and in drug discovery where it can be used to study drug transport and delivery efficacy in realistic tumor microenvironment, thereby enabling drug compound and/or delivery vehicle screening.
This paper presents a bipolar electrode (BPE) device in a microfluidic dual-channel design for co... more This paper presents a bipolar electrode (BPE) device in a microfluidic dual-channel design for concurrent preconcentration and separation of composite DNA containing samples. The novelty of the present effort relies on the combination of BPE-induced ion concentration polarization (ICP) and end-labeled free-solution electrophoresis (ELFSE). The ion concentration polarization effect arising from the faradaic reaction on the BPE is utilized to exert opposing electrophoretic and electroosmotic forces on the DNA samples. Meanwhile, end-labeled free-solution electrophoresis alters the mass-charge ratio to enable simultaneous DNA separation in free solution. The microfluidic device was fabricated using standard and soft lithography techniques to form gold-on-glass electrode capped with a PDMS microfluidic channel. Experimental testing with various DNA samples was carried out over a range of applied electric field. Concentration ratios up to 285× within 5 minutes for a 102-mer DNA, and concurrent preconcentration and free-solution separation of binary mixture of free and bound 102-mer DNA within 6 minutes was demonstrated. The effect of applied electric field was also interrogated with respect to pertinent performance metrics of preconcentration and separation.
Current in vitro models of the leukocyte adhesion cascade cannot be used for real-time studies of... more Current in vitro models of the leukocyte adhesion cascade cannot be used for real-time studies of the entire leukocyte adhesion cascade, including rolling, adhesion, and migration in a single assay. In this study, we have developed and validated a novel bioinspired microfluidic assay (bMFA) and used it to test the hypothesis that blocking of specific steps in the adhesion/migration cascade significantly affects other steps of the cascade. The bMFA consists of an endothelialized microvascular network in communication with a tissue compartment via a 3 μm porous barrier. Human neutrophils in bMFA preferentially adhered to activated human endothelial cells near bifurcations with rolling and adhesion patterns in close agreement with in vivo observations. Treating endothelial cells with monoclonal antibodies to E-selectin or ICAM-1 or treating neutrophils with wortmannin reduced rolling, adhesion, and migration of neutrophils to 60%, 20%, and 18% of their respective control values. Antibo...
This paper presents a Nafion film based micro-nanofluidic device for concurrent DNA preconcentrat... more This paper presents a Nafion film based micro-nanofluidic device for concurrent DNA preconcentration and separation. The principle of the device is based on the combination of (a) ion concentration polarization phenomenon at the junction of the microchannel and the nanochannels in the Nafion film to form opposing electrophoretic and electroosmotic forces acting on the DNAs, and (b) end-labeled free solution electrophoresis to harness the charge-to-mass ratio for molecular differentiation. The experiments successfully demonstrated concurrent preconcentration and separation of DNA mixture in free solution within 240s, yielding concentration ratios up to 1,150X and separation resolution of 1.85. The effect of applied electric field on the concentration and separation performance was also investigated. The device can be used as a key sample preparation element in conjunction with micro- or nano-fluidic sensors for microTAS functionality.
This paper presents a microfluidic electrical impedance flow cytometer (FC) for identifying the d... more This paper presents a microfluidic electrical impedance flow cytometer (FC) for identifying the differentiation state of single stem cells. This device is comprised of a novel dual micropore design, which not only enhances the processing throughput, but also allows the associated electrodes to be used as a reference for one another. A signal processing algorithm, based on the support vector machine (SVM) theory, and a data classification method were developed to automate the identification of sample types and cell differentiation state based on measured impedance values. The device itself was fabricated using a combination of standard and soft lithography techniques to generate a PDMS-gold electrode construct. Experimental testing with non-biological particles and mouse embryonic carcinoma cells (P19, undifferentiated and differentiated) was carried out using a range of excitation frequencies. The effects of the frequency and the interrogation parameters on sample identification performance were investigated. It was found that the real and imaginary part of the detected impedance signal were adequate for distinguishing the undifferentiated P19 cells from non-biological polystyrene beads at all tested frequencies. A higher frequency and an opacity index were required to resolve the undifferentiated and differentiated P19 cells by capturing capacitive changes in electrophysiological properties arising from differentiation. The experimental results demonstrated salient accuracy of the device and algorithm, and established its feasibility for non-invasive, label-free identification of the differentiation state of the stem cells.
2013 13th IEEE International Conference on Nanotechnology (IEEE-NANO 2013), 2013
ABSTRACT This paper presents a Nafion film based micro-nanofluidic device for concurrent DNA prec... more ABSTRACT This paper presents a Nafion film based micro-nanofluidic device for concurrent DNA preconcentration and separation. The principle of the device is based on: (a) ion concentration polarization phenomenon at the junction of the microchannel and the nanochannels in the Nafion film to form opposing electrophoretic and electroosmotic forces acting on the DNAs, combined with (b) end-labeled free solution electrophoresis to vary the charge-to-mass ratio for molecular differentiation. Extensive experiments were carried out to characterize the functionality of the device. Concurrent preconcentration and separation of a DNA mixture within 240s were successfully demonstrated, yielding concentration ratios up to 1,150X and separation resolution of 1.85. The effect of applied electric field on the concentration and separation performance was investigated. The device can be used as a key sample preparation element in conjunction with micro- or nanofluidic sensors to obtained microTAS functionality.
Due to the significance of the leukocyte-endothelium interactions in pathogenesis of disease and ... more Due to the significance of the leukocyte-endothelium interactions in pathogenesis of disease and drug delivery, amongst many others, several in vitro models have been developed to study different aspects of the leukocyte adhesion cascade. Current in vitro models of the leukocyte adhesion cascade cannot be used for real-time studies of the entire leukocyte adhesion cascade including rolling, adhesion and migration in a single assay. In this study, we have developed and validated a novel microfluidic assay and used it to test the hypothesis that blocking of specific steps in the adhesion/migration cascade significantly affects other steps of the cascade.
Tumor drug delivery is a complex phenomenon affected by several elements in addition to drug or d... more Tumor drug delivery is a complex phenomenon affected by several elements in addition to drug or delivery vehicle's physico-chemical properties. A key factor is tumor microvasculature with complex effects including convective transport, high interstitial pressure and enhanced vascular permeability due to the presence of "leaky vessels". Current in vitro models of the tumor microenvironment for evaluating drug delivery are oversimplified and, as a result, show poor correlation with in vivo performance. In this study, we report on the development of a novel microfluidic platform that models the tumor microenvironment more accurately, with physiologically and morphologically realistic microvasculature including endothelial cell lined leaky capillary vessels along with 3D solid tumors. Endothelial cells and 3D spheroids of cervical tumor cells were co-cultured in the networks. Drug vehicle screening was demonstrated using GFP gene delivery by different formulations of nanopolymers. The synthetic tumor network was successful in predicting in vivo delivery efficiencies of the drug vehicles. The developed assay will have critical applications both in basic research, where it can be used to develop next generation delivery vehicles, and in drug discovery where it can be used to study drug transport and delivery efficacy in realistic tumor microenvironment, thereby enabling drug compound and/or delivery vehicle screening.
This paper presents a bipolar electrode (BPE) device in a microfluidic dual-channel design for co... more This paper presents a bipolar electrode (BPE) device in a microfluidic dual-channel design for concurrent preconcentration and separation of composite DNA containing samples. The novelty of the present effort relies on the combination of BPE-induced ion concentration polarization (ICP) and end-labeled free-solution electrophoresis (ELFSE). The ion concentration polarization effect arising from the faradaic reaction on the BPE is utilized to exert opposing electrophoretic and electroosmotic forces on the DNA samples. Meanwhile, end-labeled free-solution electrophoresis alters the mass-charge ratio to enable simultaneous DNA separation in free solution. The microfluidic device was fabricated using standard and soft lithography techniques to form gold-on-glass electrode capped with a PDMS microfluidic channel. Experimental testing with various DNA samples was carried out over a range of applied electric field. Concentration ratios up to 285× within 5 minutes for a 102-mer DNA, and concurrent preconcentration and free-solution separation of binary mixture of free and bound 102-mer DNA within 6 minutes was demonstrated. The effect of applied electric field was also interrogated with respect to pertinent performance metrics of preconcentration and separation.
Current in vitro models of the leukocyte adhesion cascade cannot be used for real-time studies of... more Current in vitro models of the leukocyte adhesion cascade cannot be used for real-time studies of the entire leukocyte adhesion cascade, including rolling, adhesion, and migration in a single assay. In this study, we have developed and validated a novel bioinspired microfluidic assay (bMFA) and used it to test the hypothesis that blocking of specific steps in the adhesion/migration cascade significantly affects other steps of the cascade. The bMFA consists of an endothelialized microvascular network in communication with a tissue compartment via a 3 μm porous barrier. Human neutrophils in bMFA preferentially adhered to activated human endothelial cells near bifurcations with rolling and adhesion patterns in close agreement with in vivo observations. Treating endothelial cells with monoclonal antibodies to E-selectin or ICAM-1 or treating neutrophils with wortmannin reduced rolling, adhesion, and migration of neutrophils to 60%, 20%, and 18% of their respective control values. Antibo...
This paper presents a Nafion film based micro-nanofluidic device for concurrent DNA preconcentrat... more This paper presents a Nafion film based micro-nanofluidic device for concurrent DNA preconcentration and separation. The principle of the device is based on the combination of (a) ion concentration polarization phenomenon at the junction of the microchannel and the nanochannels in the Nafion film to form opposing electrophoretic and electroosmotic forces acting on the DNAs, and (b) end-labeled free solution electrophoresis to harness the charge-to-mass ratio for molecular differentiation. The experiments successfully demonstrated concurrent preconcentration and separation of DNA mixture in free solution within 240s, yielding concentration ratios up to 1,150X and separation resolution of 1.85. The effect of applied electric field on the concentration and separation performance was also investigated. The device can be used as a key sample preparation element in conjunction with micro- or nano-fluidic sensors for microTAS functionality.
This paper presents a microfluidic electrical impedance flow cytometer (FC) for identifying the d... more This paper presents a microfluidic electrical impedance flow cytometer (FC) for identifying the differentiation state of single stem cells. This device is comprised of a novel dual micropore design, which not only enhances the processing throughput, but also allows the associated electrodes to be used as a reference for one another. A signal processing algorithm, based on the support vector machine (SVM) theory, and a data classification method were developed to automate the identification of sample types and cell differentiation state based on measured impedance values. The device itself was fabricated using a combination of standard and soft lithography techniques to generate a PDMS-gold electrode construct. Experimental testing with non-biological particles and mouse embryonic carcinoma cells (P19, undifferentiated and differentiated) was carried out using a range of excitation frequencies. The effects of the frequency and the interrogation parameters on sample identification performance were investigated. It was found that the real and imaginary part of the detected impedance signal were adequate for distinguishing the undifferentiated P19 cells from non-biological polystyrene beads at all tested frequencies. A higher frequency and an opacity index were required to resolve the undifferentiated and differentiated P19 cells by capturing capacitive changes in electrophysiological properties arising from differentiation. The experimental results demonstrated salient accuracy of the device and algorithm, and established its feasibility for non-invasive, label-free identification of the differentiation state of the stem cells.
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Papers by Charles Garson