Using micro-patterned sensors and cell self-assembly for measuring the oxygen consumption rate of single cells

Etzkorn, James R.; Wu, Wen Chung; Zhang, Tian; Kim, Prince; Jang, Sooyoung; Meldrum, Deirdre R.; Jen, Alex K.‐Y.; Parviz, Babak A.

doi:10.1088/0960-1317/20/9/095017

Cited by 22 publications

(21 citation statements)

References 27 publications

(30 reference statements)

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“…The available experimental approaches for oxygen consumption measurements in individual cells can be divided into two groups: one set of techniques is based on microprobes, [18][19][20][21][22] whereas the second involves the isolation of individual cells in microwells. [23][24][25][26] Although microprobe methods provide a robust and sensitive tool for oxygen flux measurements at the singlecell level, they suffer from several limitations, including low throughput (one cell at a time) and the inability to separate potential contributions from adjacent cells in high-density cultures.…”

Section: Introductionmentioning

confidence: 99%

“…27 A different approach for measuring oxygen consumption rates of individual cells has been developed by our group and is based on the measurements of oxygen fluxes in isolated hermetically sealed microwells containing single cells. [23][24][25][26] In contrast to microprobe-based oxygen consumption measurements, methods based on isolation of individual cells in microwells offer at least one order of magnitude higher throughput and true single-cell measurement capability. Enclosing individual cells in hermetically sealed chambers provides a unique possibility to measure isolated oxygen consumption with true single-cell resolution.…”

Section: Introductionmentioning

confidence: 99%

“…Creating cultures of single cells in the required arrangement, i.e., individual cells placed at particular distances inside microfabricated features for the production of sealed chambers, represents a challenge in itself. The reported methods [23][24][25][26] use random cell seeding, where the bottom part (wells) of the microwells are populated with cells by randomly distributing cells suspended in media over an array of wells. Although the approach is simple and produces satisfactory results, its major limitation is the low efficiency of populating the wells with single cells.…”

Section: Introductionmentioning

confidence: 99%

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Method for physiologic phenotype characterization at the single-cell level in non-interacting and interacting cells

et al. 2012

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Abstract. Intercellular heterogeneity is a key factor in a variety of core cellular processes including proliferation, stimulus response, carcinogenesis, and drug resistance. However, cell-to-cell variability studies at the single-cell level have been hampered by the lack of enabling experimental techniques. We present a measurement platform that features the capability to quantify oxygen consumption rates of individual, non-interacting and interacting cells under normoxic and hypoxic conditions. It is based on real-time concentration measurements of metabolites of interest by means of extracellular optical sensors in cell-isolating microwells of subnanoliter volume. We present the results of a series of measurements of oxygen consumption rates (OCRs) of individual non-interacting and interacting human epithelial cells. We measured the effects of cell-to-cell interactions by using the system's capability to isolate two and three cells in a single well. The major advantages of the approach are: 1. ratiometric, intensity-based characterization of the metabolic phenotype at the single-cell level, 2. minimal invasiveness due to the distant positioning of sensors, and 3. ability to study the effects of cell-cell interactions on cellular respiration rates.

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Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Method for physiologic phenotype characterization at the single-cell level in non-interacting and interacting cells

et al. 2012

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“…SU-8 microchannels designed for continuous flows with pressure drops up to 1000kPa have been integrated with Mach-Zehnder Interferometer nanophotonic inferometric label free sensors (Blanco et al 2006). SU-8 photopatterning has also been employed to create oxygen sensor rings and cell trapping structures for the measurement of single cell oxygen consumption (Etzkorn et al 2010). Simple microfluidic devices made out paper are becoming increasingly popular.…”

Section: Microfluidic Devices Made From Photopatterned Materialsmentioning

confidence: 99%

Photopatterned materials in bioanalytical microfluidic technology

Tentori

Herr

2011

J. Micromech. Microeng.

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Microfluidic technologies are playing an increasingly important role in biological inquiry. Sophisticated approaches to the microanalysis of biological specimens rely, in part, on the fine fluid and material control offered by microtechnology, as well as a sufficient capacity for systems integration. A suite of techniques that utilize photopatterning of polymers on fluidic surfaces, within fluidic volumes, and as primary device structures underpins recent technological innovation in bioanalysis. Well-characterized photopatterning approaches enable previously fabricated or commercially fabricated devices to be customized by the user in a straight-forward manner, making the tools accessible to laboratories that do not focus on microfabrication technology innovation. In this review of recent advances, we summarize reported microfluidic devices with photopatterned structures and regions as platforms for a diverse set of biological measurements and assays.

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“…There has been an extensive need to spatially separate optical sensors and integrate them with microfeatures such as micro-channels and micro-wells [5-6]. This is especially important for multi-parameter sensing application which requires multi-spot sensor array structures.…”

Section: Introductionmentioning

confidence: 99%

Micro-patterning and characterization of PHEMA-co-PAM-based optical chemical sensors for lab-on-a-chip applications

Zhu¹,

Zhou²,

Su³

et al. 2012

Sensors and Actuators B: Chemical

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We report a novel method for wafer level, high throughput optical chemical sensor patterning, with precise control of the sensor volume and capability of producing arbitrary microscale patterns. Monomeric oxygen (O2) and pH optical probes were polymerized with 2-hydroxyethyl methacrylate (HEMA) and acrylamide (AM) to form spin-coatable and further crosslinkable polymers. A micro-patterning method based on micro-fabrication techniques (photolithography, wet chemical process and reactive ion etch) was developed to miniaturize the sensor film onto glass substrates in arbitrary sizes and shapes. The sensitivity of fabricated micro-patterns was characterized under various oxygen concentrations and pH values. The process for spatially integration of two sensors (Oxygen and pH) on the same substrate surface was also developed, and preliminary fabrication and characterization results were presented. To the best of our knowledge, it is the first time that poly (2-hydroxylethyl methacrylate)-co-poly (acrylamide) (PHEMA-co-PAM)-based sensors had been patterned and integrated at the wafer level with micron scale precision control using microfabrication techniques. The developed methods can provide a feasible way to miniaturize and integrate the optical chemical sensor system and can be applied to any lab-on-a-chip system, especially the biological micro-systems requiring optical sensing of single or multiple analytes.

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Using micro-patterned sensors and cell self-assembly for measuring the oxygen consumption rate of single cells

Cited by 22 publications

References 27 publications

Method for physiologic phenotype characterization at the single-cell level in non-interacting and interacting cells

Method for physiologic phenotype characterization at the single-cell level in non-interacting and interacting cells

Photopatterned materials in bioanalytical microfluidic technology

Micro-patterning and characterization of PHEMA-co-PAM-based optical chemical sensors for lab-on-a-chip applications

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