Toward the Analysis of Mitochondria Isolated from Leukemic Cells with Electrochemically Instrumented Microwell Arrays

Lemercier, Gabriel; Sekli-Belaidi, Fadhila; Vajrala, Suresh; Descamps, Emeline; Foncy, Julie; Arbault, Stéphane; Sarry, Jean-Emmanuel; Temple‐Boyer, Pierre; Launay, Jérôme

doi:10.3390/proceedings1040289

Cited by 2 publications

(3 citation statements)

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“…In Equation (3), it should be noticed that

C_{O 2}^{cell}

is representative of the cellular oxygen consumption and it depends on the glucose concentration in solution. [ 39,40 ] Since the dissolved oxygen reductive current i RNE measured on the Pt ring nanoelectrode is proportional to

C_{O 2}^{RNE}

and the OptoElecWell device was designed to prevent crosstalk between microwells, [ 30,41 ] the measured currents should follow temporal variations according to a “Cottrell‐like relation.” [ 42 ]

\begin{matrix} (- i_{measured}) = (- i_{final}) + \frac{K}{\sqrt t} \end{matrix}

Where the final current i final and the parameter K , directly proportional to the oxygen consumption rate, depends on the glucose concentration in solution.…”

Section: Methodsmentioning

confidence: 99%

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Microwell Array Based Opto‐Electrochemical Detections Revealing Co‐Adaptation of Rheological Properties and Oxygen Metabolism in Budding Yeast

et al. 2021

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Microdevices composed of microwell arrays integrating nanoelectrodes (OptoElecWell) are developed to achieve dual high‐resolution optical and electrochemical detections on single Saccharomyces cerevisiae yeast cells. Each array consists of 1.6 × 105 microwells measuring 8 µm in diameter and 5 µm height, with a platinum nanoring electrode for in situ electrochemistry, all integrated on a transparent thin wafer for further high‐resolution live‐cell imaging. After optimizing the filling rate, 32% of cells are effectively trapped within microwells. This allows to analyse S. cerevisiae metabolism associated with basal respiration while simultaneously measuring optically other cellular parameters. In this study, the impact of glucose concentration on respiration and intracellular rheology is focused. It is found that while the oxygen uptake rate decreases with increasing glucose concentration, diffusion of tracer nanoparticles increases. The OptoElecWell‐based respiration methodology provides similar results compared to the commercial gold‐standard Seahorse XF analyzer, while using 20 times fewer biological samples, paving the way to achieve single cell metabolomics. In addition, it facilitates an optical route to monitor the contents within single cells. The proposed device, in combination with the dual detection analysis, opens up new avenues for measuring cellular metabolism, and relating it to cellular physiological indicators at single cell level.

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“…In Equation (3), it should be noticed that

C_{O 2}^{cell}

C_{O 2}^{RNE}

\begin{matrix} (- i_{measured}) = (- i_{final}) + \frac{K}{\sqrt t} \end{matrix}

Where the final current i final and the parameter K , directly proportional to the oxygen consumption rate, depends on the glucose concentration in solution.…”

Section: Methodsmentioning

confidence: 99%

“…and the OptoElecWell device was designed to prevent crosstalk between microwells, [30,41] the measured currents should follow temporal variations according to a "Cottrell-like relation." [42] measured final ( ) ( )…”

mentioning

confidence: 99%

Microwell Array Based Opto‐Electrochemical Detections Revealing Co‐Adaptation of Rheological Properties and Oxygen Metabolism in Budding Yeast

et al. 2021

Self Cite

View full text Add to dashboard Cite

show abstract

“…In equation 3, it should be noticed that 2 cell is representative of the cellular oxygen consumption and it depends on the glucose concentration in solution (Barnabé and Butler, 2000;Takeda et al, 2015). Since the dissolved oxygen reductive current RNE measured on the Pt ring nanoelectrode is proportional to 2 and the OptoElecWell device was designed to prevent crosstalk between microwells (Lemercier et al, 2017;Sékli Belaïdi et al, 2016), the measured currents should follow temporal variations according to a "Cottrell-like relation" (A.J. Bard and L R Faulkner, 2000).…”

Section: Model For Oxygen Consumption Rate Monitoring Within Optoelecmentioning

confidence: 99%

Microwell array based opto-electrochemical detections revealing co-adaptation of rheological properties and oxygen metabolism in budding yeast

Vajrala

Alric

Laborde

et al. 2021

Preprint

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Microdevices composed of microwell arrays integrating nanoelectrodes (OptoElecWell) were developed to achieve dual high-resolution optical and electrochemical detections on single Saccharomyces cerevisiae budding yeast cells. Each array consists in 1.6 × 105 microwells of 8 µm diameter and 5 µm height, with a platinum nanoring electrode for in-situ electrochemistry, all integrated on a transparent thin wafer for further high-resolution live-cell imaging. After optimizing the filling rate, 32% of cells were effectively trapped within microwells. This allowed to analyse S. cerevisiae metabolisms associated with basal respiration while simultaneously measuring optically other cellular parameters. In this study, we focused on the impact of glucose concentration on respiration and intracellular rheology. We found that while oxygen uptake rate decreased with increasing glucose concentration, diffusion of tracer nanoparticles increased. Our OptoElecWell based respiration methodology provided similar results compared to the commercial gold-standard Seahorse XF analyser, while using 20 times lesser biological samples, paving the way to achieve single cell metabolomics. In addition, it facilitates an optical route to monitor the contents within single cells. The proposed device, in combination with the dual detection analysis, opens up new avenues for measuring cellular metabolism, and relating it to various cellular physiological and rheological indicators at single cell level.

show abstract

Toward the Analysis of Mitochondria Isolated from Leukemic Cells with Electrochemically Instrumented Microwell Arrays

Cited by 2 publications

References 4 publications

Microwell Array Based Opto‐Electrochemical Detections Revealing Co‐Adaptation of Rheological Properties and Oxygen Metabolism in Budding Yeast

Microwell Array Based Opto‐Electrochemical Detections Revealing Co‐Adaptation of Rheological Properties and Oxygen Metabolism in Budding Yeast

Microwell array based opto-electrochemical detections revealing co-adaptation of rheological properties and oxygen metabolism in budding yeast

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