A printed superoxide dismutase coated electrode for the study of macrophage oxidative burst

Hiatt, Leslie A.; McKenzie, Jennifer R.; Deravi, Leila F.; Harry, Reese S.; Wright, David W.; Cliffel, David E.

doi:10.1016/j.bios.2011.12.038

Cited by 15 publications

(17 citation statements)

References 48 publications

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“…26 While these devices allowed for real-time detection of metabolites, they were difficult to seal without crushing the cells or blocking the electrodes with PDMS or bubbles, limiting reproducibility. For this work, two custom acrylic housings were designed to house either a single SPE or a six-sensor array of SPEs.…”

Section: Methodsmentioning

confidence: 99%

Real-Time Monitoring of Cellular Bioenergetics with a Multianalyte Screen-Printed Electrode

et al. 2015

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Real-time monitoring of changes to cellular bioenergetics can provide new insights into mechanisms of action for disease and toxicity. This work describes the development of a multi-analyte screen-printed electrode for the detection of analytes central to cellular bioenergetics: glucose, lactate, oxygen, and pH. Platinum screen-printed electrodes were designed in-house and printed by Pine Research Instrumentation. Electrochemical plating techniques were used to form quasi-reference and pH electrodes. A Dimatix materials inkjet printer was used to deposit enzyme and polymer films to form sensors for glucose, lactate, and oxygen. These sensors were evaluated in bulk solution and microfluidic environments, and found to behave reproducibly and possess a lifetime of up to six weeks. Linear ranges and limits of detection for enzyme-based sensors were found to have an inverse relationship with enzyme loading, and iridium oxide pH sensors were found to have super-Nernstian responses. Preliminary measurements where the sensor was enclosed within a microfluidic channel with RAW 264.7 macrophages were performed to demonstrate the sensors’ capabilities for performing real-time microphysiometry measurements.

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Section: Methodsmentioning

confidence: 99%

Real-Time Monitoring of Cellular Bioenergetics with a Multianalyte Screen-Printed Electrode

et al. 2015

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“…Although chemical measurements can be performed off-chip [7], integrating analytical measurements with cells on microfluidic systems eliminates the need for sample collection and off-line analyte detection, and can provide real-time recording of cellular dynamics [8,9]. Several examples of this approach have been described including on-line immunoassays [10], sensors [11], and enzyme assays [12]. Most such systems perform a single assay on cells.…”

Section: Introductionmentioning

confidence: 99%

Multiplexed microfluidic enzyme assays for simultaneous detection of lipolysis products from adipocytes

et al. 2014

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Microfluidics have enabled new cell biology experiments. Incorporating chemical monitoring of cellular secretion into chips offers the potential to increase information content and utility of such systems. In this work, an integrated, multilayer polydimethylsiloxane microfluidic chip was developed to simultaneously measure fatty acids and glycerol secreted from cultured adipocytes on-chip in near real-time. Approximately 48,000 adipocytes were loaded into a cell chamber in a reversibly-sealed chip. Cells were perfused at 0.75 μL/min. Cell perfusate was split and directed to separate, continuously operating fluorescent enzyme assay channel networks. The fluorescent assay products were detected simultaneously near the outlet of the chip. The fatty acid and glycerol assays had linear dynamic ranges to 150 μM and 110 μM, and LOD of 6 μM and 5 μM, respectively. Surface modifications including pretreatment with sodium dodecyl sulfate were utilized to prevent adsorption of fatty acids to the chip surface. Using the chip, basal fatty acid and glycerol concentrations ranged from 0.18–0.7 nmol 106 cell−1 min−1 and 0.23–0.85 nmol 106 cell−1 min−1, respectively. Using valves built into the chip, the perfusion solution was switched to add 20 μM isoproterenol, a β-adrenergic agonist, which stimulates the release of glycerol and fatty acids in adipocytes. This manipulation resulted in a rapid and stable 1.5- to 6.0-fold increase of NEFA and glycerol. The ratio of NEFA to glycerol released increased with adipocyte age. These experiments illustrate the potential for performing multiple real-time assays on cells in culture using microfluidic devices.

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“…For this configuration, a screen-printed multianalyte sensor (SPE) was designed in-house and printed by Pine Research Instrumentation [16]. One electrode on the SPE was plated with a pH-sensitive IrOx film [28].…”

Section: Instrumentsmentioning

confidence: 99%

“…This also allowed for expansion of the MAMP technique to other platforms [16]. In this configuration, two multipotentiostat modules are replaced with two OCP modules, each containing three independent channels.…”

Section: Introductionmentioning

confidence: 99%

Multichamber multipotentiostat system for cellular microphysiometry

Lima

Snider

Reiserer

et al. 2014

Sensors and Actuators B: Chemical

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Multianalyte microphysiometry is a powerful technique for studying cellular metabolic flux in real time. Monitoring several analytes concurrently in a number of individual chambers, however, requires specific instrumentation that is not available commercially in a single, compact, benchtop form at an affordable cost. We developed a multipotentiostat system capable of performing simultaneous amperometric and potentiometric measurements in up to eight individual chambers. The modular design and custom LabVIEW™ control software provide flexibility and allow for expansion and modification to suit different experimental conditions. Superior accuracy is achieved when operating the instrument in a standalone configuration; however, measurements performed in conjunction with a previously developed multianalyte microphysiometer have shown low levels of crosstalk as well. Calibrations and experiments with primary and immortalized cell cultures demonstrate the performance of the instrument and its capabilities.

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A printed superoxide dismutase coated electrode for the study of macrophage oxidative burst

Cited by 15 publications

References 48 publications

Real-Time Monitoring of Cellular Bioenergetics with a Multianalyte Screen-Printed Electrode

Real-Time Monitoring of Cellular Bioenergetics with a Multianalyte Screen-Printed Electrode

Multiplexed microfluidic enzyme assays for simultaneous detection of lipolysis products from adipocytes

Multichamber multipotentiostat system for cellular microphysiometry

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