Biomedical subjects

A Bruno Frazier

Publications and source records attributed to A Bruno Frazier.

10 recordsLinked to original sources

PubMed2007

Quantification of the heterogeneity in breast cancer cell lines using whole-cell impedance spectroscopy.

PURPOSE: Quantification of the heterogeneity of tumor cell populations is of interest for many diagnostic and therapeutic applications, including determining the cancerous stage of tumors. We attempted to differentiate human breast cancer cell lines from different pathologic stages and compare that with a normal human breast tissue cell line by characterizing the impedance properties of each cell line. EXPERIMENTAL DESIGN: A microelectrical impedance spectroscopy system has been developed that can trap a single cell into an analysis cavity and measure the electrical impedance of the captured cell over a frequency range from 100 Hz to 3.0 MHz. Normal human breast tissue cell line MCF-10A, early-stage breast cancer cell line MCF-7, invasive human breast cancer cell line MDA-MB-231, and metastasized human breast cancer cell line MDA-MB-435 were used. RESULTS: The whole-cell impedance signatures show a clear difference between each cell line in both magnitude and phase of the electrical impedance. The membrane capacitance calculated from the impedance data was 1.94 +/- 0.14, 1.86 +/- 0.11, 1.63 +/- 0.17, and 1.57 +/- 0.12 muF/cm(2) at 100 kHz for MCF-10A, MCF-7, MDA-MB-231, and MDA-MB-435, respectively. The calculated resistance for each cancer cell line at 100 kHz was 24.8 +/- 1.05, 24.8 +/- 0.93, 24.9 +/- 1.12, and 26.2 +/- 1.07 MOhm, respectively. The decrease in capacitances of the cancer cell lines compared with that of the normal cell line MCF-10A was 4.1%, 16.0%, and 19.1%, respectively, at 100 kHz. CONCLUSIONS: These findings suggest that microelectrical impedance spectroscopy might find application as a method for quantifying progression of cancer cells without the need for tagging or modifying the sampled cells.

Breast Neoplasms↗

PubMed2007

Three dimensional MEMS microfluidic perfusion system for thick brain slice cultures.

In vitro tissue culture models are often benchmarked by their ability to replicate in vivo function. One of the limitations of in vitro systems is the difficulty in preserving an orchestrated cell population, especially for generating three-dimensional tissue equivalents. For example, tissue-engineering applications involve large high-density constructs, requiring a perfusing system that is able to apply adequate oxygen and nutrients to the interior region of the tissue. This is particularly true with respect to thick tissue sections harvested for in vitro culture. We have fabricated a microneedle-based perfusion device for high-cell-density in vitro tissue culture from SU-8 photosensitive epoxy and suitable post-processing. The device was tested for its ability to improve viability in slices of harvested brain tissue. This model was chosen due to its acute sensitivity to disruptions in its nutrient supply. Improved viability was visible in the short term as assessed via live-dead discriminating fluorescent staining and confocal microscopy. This perfusion system opens up many possibilities for both neurobiological as well as other culture systems.

Animals↗

PubMed2006

A multielectrode microcompartment culture platform for studying signal transduction in the nervous system.

This paper describes the design, fabrication, and characterization of a microfabricated compartmented culture system (micro-CCS) useful for electrophysiological signaling studies in cultured neurons. The focus of the paper is the process of interfacing the micro-CCS with cultured neurons and to demonstrate the applicability of the system for biochemical-mediated electrophysiological studies. Moreover, we show that we can record action potentials from cultured neurons through the extracellular compartmented application of elevated levels of K(+) ions. Finally, we show that we can isolate the electrophysiological effects of the sodium channel blocker tetrodotoxin in one of the compartments of a two compartment culture while recording electrophysiological data from both compartments.

Animals↗

PubMed2006

Ion channel characterization using single cell impedance spectroscopy.

A micro electrical impedance spectroscopy system (microEIS) for single cell analysis has been developed and used to differentiate ion channel activities of bovine chromaffin cells. K+ and Ca2+ channels were blocked and their electrical impedances were measured over a frequency range of 100 Hz to 5.0 MHz and compared to that of unblocked chromaffin cells. When ion channels were blocked, an increase in magnitude and decrease in phase of the measured impedances were observed. This result demonstrates that ion channel activities can be distinguished using the developed microsystem and it is expected that this system can be used to provide positive/negative information of ion channel blockage in a high throughput screening setup.

Animals↗

PubMed2006

A compartmented neuronal culture system in microdevice format.

This paper describes a microfabricated compartmented culture system (mu-CCS) for studying the effects of drugs on cultured neurons. We describe the fabrication of the microsystem and show the ability to culture DRG neurons in the microsystem. Furthermore, we demonstrate the ability to culture neurons with extensions growing into adjoining compartments while maintaining fluid isolation between compartments. The axonal growth pattern was controlled along the surface of the glass microelectrode substrate using a micropatterned collagen film. The ability to isolate fluids to selected compartments while simultaneously allowing intercompartmental growth of the axons enables various studies in which selected segments of neurons or populations of neurons can be selectively exposed to biochemical treatment. The neurotoxin vincristine was used as the test vehicle to demonstrate the functionality of the mu-CCS. Vincristine was applied to the axonal compartment to show that the interaction of drugs with DRG neurons progresses in a way similar to neurons cultured/exposed using conventional techniques.

Animals↗

PubMed2006

Microsystems for isolation and electrophysiological analysis of breast cancer cells from blood.

This paper presents the development of a microsystem for separating suspended breast cancer cells in peripheral blood and for sorting them based on their electrophysiological characteristics. A continuous paramagnetic capture mode (PMC) magnetophoretic microseparator was utilized for the isolation of suspended breast cancer cells in peripheral blood based on the native magnetic properties of blood cells without any tagging such as with magnetic probes. A micro-electrical impedance spectroscopy (mu-EIS) system was used as a downstream cell analysis tool to extract the pathological characteristics from the breast cancer cells. The system was fabricated on silicon and glass substrates utilizing microfabrication and stereolithography technologies. The experimental results of the PMC microseparator show that 94.8% of the breast cancer cells could be continuously separated out from a spiked blood sample with a 0.2 T external magnetic flux. The electrical impedances of human breast cancer cell lines of different pathological stages (MCF-7, MDA-MB-231, and MDA-MB-435) were measured using mu-EIS and compared to those of normal human breast tissue cell line MCF-10A.

Breast Neoplasms↗

PubMed2005

Paramagnetic capture mode magnetophoretic microseparator for high efficiency blood cell separations.

This paper presents the characterization of continuous single-stage and three-stage cascade paramagnetic capture (PMC) mode magnetophoretic microseparators for high efficiency separation of red and white blood cells from diluted whole blood based on their native magnetic properties. The separation mechanism for both PMC microseparators is based on a high gradient magnetic separation (HGMS) method. This approach enables separation of blood cells without the use of additives such as magnetic beads. Experimental results for the single-stage PMC microseparator show that 91.1% of red blood cells were continuously separated from the sample at a volumetric flow rate of 5 microl h-1. In addition, the three-stage cascade PMC microseparator continuously separated 93.5% of red blood cells and 97.4% of white blood cells from whole blood at a volumetric flow rate of 5 microl h-1.

Algorithms↗

PubMed2003

Multi-layer plastic/glass microfluidic systems containing electrical and mechanical functionality.

This paper describes an approach for fabricating multi-layer microfluidic systems from a combination of glass and plastic materials. Methods and characterization results for the microfabrication technologies underlying the process flow are presented. The approach is used to fabricate and characterize multi-layer plastic/glass microfluidic systems containing electrical and mechanical functionality. Hot embossing, heat staking of plastics, injection molding, microstenciling of electrodes, and stereolithography were combined with conventional MEMS fabrication techniques to realize the multi-layer systems. The approach enabled the integration of multiple plastic/glass materials into a single monolithic system, provided a solution for the integration of electrical functionality throughout the system, provided a mechanism for the inclusion of microactuators such as micropumps/valves, and provided an interconnect technology for interfacing fluids and electrical components between the micro system and the macro world.

Biocompatible Materials↗

PubMed2002

A microfabricated thermal field-flow fractionation system.

A microscale thermal field-flow fractionation (micro-TFFF) system has been designed, fabricated, and characterized. Motivation for miniaturization of TFFF systems was established by examining the geometrical scaling of the fundamental TFFF theory. Miniaturization of conventional macroscale TFFF systems was made possible through utilization of micromachining technologies. Fabrication of the micro-TFFF system was discussed in detail. The micro-TFFF system was characterized for plate height versus flow rate, single-component polystyrene retention, and multicomponent polystyrene separations. Retention, thermal diffusion coefficients, and maximum diameter-based selectivity values were extracted from separation data and found comparable with macroscale TFFF system results. Retention values ranged from 0.33 to 0.46. Thermal diffusion coefficients were between 3.0 x 10(-8) and 5.4 x 10(-8) cm2/s x K. The maximum diameter-based selectivity was 1.40.

Journal Article↗

PubMed2002

Geometric scaling effects in electrical field flow fractionation. 2. Experimental results.

Geometric scaling of microelectrical field flow fractionation (micro-EFFF) systems is investigated experimentally and compared to theory and to macroscale EFFF systems. Experimental results are presented to demonstrate that the miniaturized system operates according to the scaling theory associated with the system. Demonstrated improvements in the channels include increased retention and resolution and decreased peak broadening, electrical time constants, relaxation time, power consumption, and sample size. Additionally, scaling effects related to the compression of separation zones in the miniaturized EFFF systems are discussed.

Electrodes↗