Metabolic oxygen consumption measurement with a single-cell biosensor after particle microbeam irradiation

Xu, Yanping; Zhang, Bo; Messerli, Mark A.; Randers-Pehrson, Gerhard; Hei, Tom K.; Brenner, David J.

doi:10.1007/s00411-014-0574-1

Cited by 3 publications

(3 citation statements)

References 9 publications

(9 reference statements)

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“…For radiobiology studies, ion microbeam systems [9, 10 and 11] that have been developed in the last decade allow the irradiation of small structures such as the nuclei of cultured cells growing in a petri dish using a beam of energetic ions produced in an accelerator and then confined to a few micron diameter. A typical experiment is to allow one or a few ions to pass through each target that has been located and placed in the path of the beam [12]. However, the current developed horizontal microbeam is aiming to produce a novel secondary particle microbeam (neutron microbeam) or to conduct a microanalysis (microPIXE) which all require a high initial ion beam (proton beam mostly) current.…”

Section: Discussionmentioning

confidence: 99%

A horizontal multi-purpose microbeam system

Randers-Pehrson

Marino

et al. 2018

Nuclear Instruments and Methods in Physics Research Section A:

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A horizontal multi-purpose microbeam system with a single electrostatic quadruplet focusing lens has been developed at the Columbia University Radiological Research Accelerator Facility (RARAF). It is coupled with the RARAF 5.5 MV Singleton accelerator (High Voltage Engineering Europa, the Netherlands) and provides micrometer-size beam for single cell irradiation experiments. It is also used as the primary beam for a neutron microbeam and microPIXE (particle induced x-ray emission) experiment because of its high particle fluence. The optimization of this microbeam has been investigated with ray tracing simulations and the beam spot size has been verified by different measurements.

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Section: Discussionmentioning

confidence: 99%

A horizontal multi-purpose microbeam system

Randers-Pehrson

Marino

et al. 2018

Nuclear Instruments and Methods in Physics Research Section A:

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“…However, the technologies available before OxySplots have generally been inconvenient and in some cases cumbersome. Polarization-electrodes like the classic Clark electrode [10] or fiber-optic electrodes [12] require specialized or customized fabricated instrumentation and may interfere with patch clamp hardware or micro-tools used in mechanotransduction experiments. Other intriguing technologies include "nanobead" pO 2 sensors.…”

Section: Other Existing Technologies For Measurements Of Po 2 In Single Cells Experimentsmentioning

confidence: 99%

“…There are two primary problems for single cell experiments: (1) the large size and (2) the slow response of the Clark-type electrodes. More recently custom fabrication of miniaturized Clark-type electrode system was described [10] and additional industrial and scientific sensors have been developed. This includes the solid-state zirconium dioxide O 2 sensor [11], or a proprietary fiber optic method by Ocean Optics that has improved response time and diverse uses [12].…”

Section: Introductionmentioning

confidence: 99%

Real-time local oxygen measurements for high resolution cellular imaging

Boyman

Williams

Wescott

et al. 2019

Journal of Molecular and Cellular Cardiology

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Single-cell metabolic investigations are hampered by the absence of flexible tools to measure local partial pressure of O 2 (pO 2 ) at high spatial-temporal resolution. To this end, we developed an optical sensor capable of measuring local pericellular pO 2 for subcellular resolution measurements with confocal imaging while simultaneously carrying out electrophysiological and/or chemomechanical single cell experiments. Here we present the OxySplot optrode, a ratiometric fluorescent O 2 -micro-sensor created by adsorbing O 2 -sensitive and O 2 -insensitive fluorophores onto micro-particles of silica. To protect the OxySplot optrode from the components and reactants of liquid environment without compromising access to O 2 , the microparticles are coated with an optically clear silicone polymer (PDMS, polydimethylsiloxane). The PDMS coated OxySplot micro-particles are used alone or in a thin (~50 micron) PDMS layer of arbitrary shape referred to as the OxyMat. Additional top coatings on the OxyMat (e.g., fibronectin, laminin, polylysine, special photoactivatable surfaces etc.) facilitate adherence of cells. The OxySplots report the cellular pO 2 and micro-gradients of pO 2 without disrupting the flow of extracellular solutions or interfering with patch-clamp pipettes, mechanical attachments, and micro-superfusion. Since OxySplots and a cell can be imaged and spatially resolved, calibrated changes of pO 2 and intracellular events can be imaged simultaneously. In addition, the response-time (t 0.5 = 0.7 s, 0 -160 mm Hg) of OxySplots is ~100 times faster than amperometric Clark-type polarization microelectrodes. Two usage example of OxySplots with cardiomyocytes show (1) OxySplots measuring pericellular pO 2 while tetramethylrhodamine methyl-ester (TMRM) was used to measure mitochondrial membrane potential (ΔΨ m ); and (2) OxySplots measuring pO 2 during ischemia and reperfusion while rhod-2 was used to measure cytosolic [Ca 2+ ] i levels

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Advances in Electrochemistry for Monitoring Cellular Chemical Flux

Sarkar

2019

CMC

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The transport of molecules and inorganic ions across the plasma membrane results in chemical fluxes that reflect cellular function in healthy and diseased states. Measurement of these chemical fluxes enables the characterization of protein function and transporter stoichiometry, characterization of the viability of single cells and embryos prior to implantation, and screening of pharmaceutical agents. Electrochemical sensors are sensitive and noninvasive tools for measuring chemical fluxes immediately outside the cells in the boundary layer, that are capable of monitoring a diverse range of transported analytes including inorganic ions, gases, neurotransmitters, hormones, and pharmaceutical agents. Used on their own or in combination with other methods, these sensors continue to expand our understanding of the function of rare cells and small tissues. Advances in sensor construction and detection strategies continue to improve sensitivity under physiological conditions, diversify analyte detection, and increase throughput. These advances will be discussed in the context of addressing technical challenges to measuring in the boundary layer of cells and measuring the resultant changes to the chemical concentration in the bulk media.

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Metabolic oxygen consumption measurement with a single-cell biosensor after particle microbeam irradiation

Cited by 3 publications

References 9 publications

A horizontal multi-purpose microbeam system

A horizontal multi-purpose microbeam system

Real-time local oxygen measurements for high resolution cellular imaging

Advances in Electrochemistry for Monitoring Cellular Chemical Flux

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