A Clark-type oxygen chip for in situ estimation of the respiratory activity of adhering cells

Wu, Ching-Chou; Luk, Hsiang‐Ning; Lin, Yen-Ting; Yuan, Chia-Yin

doi:10.1016/j.talanta.2009.11.062

Cited by 52 publications

(46 citation statements)

References 36 publications

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“…The average residual currents for oxygen depleted solution measured in oxygen depleted solution were 3.4%, 1.0%, 1.0% and 2.0% of the current measured in oxygen saturated solution for flow rate 0.0, 0.2, 0.5, 1.0 ml/min respectively. The residual currents measured in this experiment were comparable to that of the Wu et al' works [31], Park's report [17,32] and Suzuki's studies [33][34][35]. The reduction currents in DPBS were also measured to verify the calibration curve, and the data were also fitted well into the calibration curve under different flow rates.…”

Section: Calibration Curvesupporting

confidence: 85%

A low temperature co-fired ceramic based microfluidic Clark-type oxygen sensor for real-time oxygen sensing

Luo

Dziubla

Eitel

2017

Sensors and Actuators B: Chemical

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a b s t r a c tIn this paper, a microfluidic platform for real-time monitoring of dissolved oxygen in a flowing microfluidic environment fabricated using low temperature co-fired ceramic (LTCC) technology is described. The fabricated Clark-type oxygen sensor consisted of three electrodes (working electrode, counter electrode and Ag/AgCl reference electrode), a solid-state proton conductive matrix (Nafion 117 membrane) and polydimethylosiloxane (PDMS) as the oxygen permeable membrane (OPM). The use of a solid-state proton conductive matrix as the electrolyte in the design of the oxygen sensor makes it feasible integrate this device in a typical LTCC fabrication process. Cyclic voltammetry and chronoamperometry measurement were used to characterize electrochemical properties of the developed oxygen sensor. The reduction current was linearly related with the dissolved oxygen concentration ranging from 0 to 8.1 mg/l under different flow conditions (0.0-1.0 ml/min). The residual currents of the oxygen sensor were less than 3.5% of that measured in oxygen saturated state, and the average response time was 10.9 s. The current device represents an improved Clark-type oxygen sensor with the advantages of easy fabrication, flexible configuration, fast response time, incorporation of microfluidic analyte introduction and real-time detection of dissolved oxygen. The potential applications include material synthesis, cell culture, biological assays incorporating controlled introduction of reagents or analytes and real-time monitoring of dissolved oxygen in a microfluidic environment.

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Section: Calibration Curvesupporting

confidence: 85%

A low temperature co-fired ceramic based microfluidic Clark-type oxygen sensor for real-time oxygen sensing

Luo

Dziubla

Eitel

2017

Sensors and Actuators B: Chemical

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“…17 Detection techniques such as the Whalen-style platinum electrode 18 allow for the measurement of O 2 consumption at the single cell level, 19 but only deliver data for one cell at a time. Other optical intracellular oxygen sensing probes and techniques including Clark-type oxygen chips 20, 21 often require highly specialized equipment such as fluorescence life time microscopy technologies. 21, 22 As many laboratories routinely use confocal or epifluorescence time-lapse imaging, there is a significant need for the development of probes for these applications.…”

mentioning

confidence: 99%

Single-cell time-lapse imaging of intracellular O2 in response to metabolic inhibition and mitochondrial cytochrome-c release

et al. 2017

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The detection of intracellular molecular oxygen (O2) levels is important for understanding cell physiology, cell death, and drug effects, and has recently been improved with the development of oxygen-sensitive probes that are compatible with live cell time-lapse microscopy. We here provide a protocol for the use of the nanoparticle probe MitoImage-MM2 to monitor intracellular oxygen levels by confocal microscopy under baseline conditions, in response to mitochondrial toxins, and following mitochondrial cytochrome-c release. We demonstrate that the MitoImage-MM2 probe, which embeds Pt(II)-5,10,15,20-tetrakis-(2,3,4,5,6–pentafluorophenyl)-porphyrin as oxygen sensor and poly(9,9-dioctylfluorene) as an O2-independent component, enables quantitative, ratiometric time-lapse imaging of intracellular O2. Multiplexing with tetra-methyl-rhodamine-methyl ester in HeLa cervical cancer cells showed significant increases in intracellular O2 accompanied by strong mitochondrial depolarization when respiratory chain complexes III or IV were inhibited by Antimycin A or sodium azide, respectively, and when cells were maintained at ‘physiological’ tissue O2 levels (5% O2). Multiplexing also allowed us to monitor intracellular O2 during the apoptotic signaling process of mitochondrial outer membrane permeabilization in HeLa expressing cytochrome-c-eGFP, and demonstrated that mitochondria post cytochrome-c release are able to retain their capacity to respire at physiological O2 despite a decrease in mitochondrial membrane potential.

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“…To measure the oxygen consumption rates (OCR) of single cells, some methodologies have been developed either using amperometric electrochemical sensing [3][4] or luminescent optical sensing [5][6][7]. In this study, we proposed a digital light modulation system that utilizes a modified commercial DMD projector equipped with a UV LED as a light modulation source to modulate the excitation light in the spatial and temporal domains.…”

Section: Introductionmentioning

confidence: 99%

Light-addressable measurements of cellular oxygen consumption rates in microwell arrays based on phase-based phosphorescence lifetime detection

Chu

Lin

Hung

et al. 2013

2013 Transducers &Amp; Eurosensors XXVII: The 17th International Conference on Solid-State Sensors, Actuators and Microsystems

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A digital light modulation system that utilizes a modified commercial digital micromirror device (DMD) projector, which is equipped with a UV LED as a light modulation source, has been developed to spatially direct excited light toward a microwell array device to detect the oxygen consumption rate (OCR) of single cells via phase-based phosphorescence lifetime detection. The microwell array device is composed of a combination of two components: an array of glass microwells containing Pt(II) octaethylporphine (PtOEP) as the oxygen-sensitive luminescent layer and a microfluidic module with pneumatically actuated glass lids set above the microwells to controllably seal the microwells of interest. By controlling the illumination pattern on the DMD, the modulated excitation light can be spatially projected to only excite the sealed microwell for cellular OCR measurements. The OCR of BHK-21 fibroblast cells cultivated on the PtOEP layer within a sealed microwell has been successfully measured at 104 ± 2.96 amol s -1 cell -1 . Repeatable and consistent measurements indicate that the oxygen measurements did not adversely affect the physiological state of the measured cells. The OCR of the cells exhibited a good linear relationship with the diameter of the microwells, ranging from 400 to 1000 m and containing approximately 480 to 1200 cells within a microwell. In addition, the OCR variation of single cells in situ infected by Dengue virus with a different multiplicity of infection (M.O.I) was also successfully measured in real-time. This proposed platform provides the potential for a wide range of biological applications in cell-based biosensing, toxicology, and drug discovery.

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A Clark-type oxygen chip for in situ estimation of the respiratory activity of adhering cells

Cited by 52 publications

References 36 publications

A low temperature co-fired ceramic based microfluidic Clark-type oxygen sensor for real-time oxygen sensing

A low temperature co-fired ceramic based microfluidic Clark-type oxygen sensor for real-time oxygen sensing

Single-cell time-lapse imaging of intracellular O2 in response to metabolic inhibition and mitochondrial cytochrome-c release

Light-addressable measurements of cellular oxygen consumption rates in microwell arrays based on phase-based phosphorescence lifetime detection

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