Fluorescence oxygen sensor using photo quenching with oxygen molecule

Aizawa, Hiroaki; Katsumata, T.; Komuro, Shuji

doi:10.1109/sice.2007.4421269

“…Comparing to the existing method of using morphological assessment for oocyte/embryo quality [11], the ability to measure mitochondrial respiration of single oocyte under different stimuli will enable better understanding of oocyte and embryo physiology and provide metrics for better prediction of oocyte/embryo quality for applications, such as assisted reproduction technologies [9]. The use of oxygen sensors for in-vitro monitoring of cell activities and for determining the effect of drugs on cell metabolism has been demonstrated using optical techniques such as florescence quenching by measuring the change in oxygen concentration near the cell [12,13]. This method has drawbacks such as temperature sensitivity, and it requires a long time to reach stability [13,14].…”

Section: Introductionmentioning

confidence: 99%

Monitoring oocyte/embryo respiration using electrochemical-based oxygen sensors

Obeidat

¹

,

Evans

²

,

Tedjo

³

et al. 2018

Sensors and Actuators B: Chemical

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Current commercially available instruments for monitoring mitochondrial respiration are incapable of single cell measurements. Therefore, we developed a three-electrode, Clark-type biosensor suitable for mitochondrial respirometry in single oocytes and embryos. The biosensor was embedded in a PMMA (polymethyl methacrylate) micro-chamber to allow investigation of single oocytes/embryos immersed in up to 100 µL of respiration buffer. The micro-chamber was completely sealed to avoid oxygen exchange between the inside of the chamber and the atmosphere, while being maintained at a temperature of 38.5 ˚C to preserve cell viability. Using amperometry, the oxygen consumption of cells inside the micro-chamber was measured as a change in output current and converted to femto-mol (fmol) oxygen consumed per second based on calibrations with known buffer oxygen concentrations. The sensor measured basal cell respiration supported by endogenous substrates, respiration associated with proton leak induced by inhibition of the adenosine triphosphate (ATP) synthase (complex V) with oligomycin, and the maximal noncoupled respiratory capacity revealed by Carbonyl cyanide-4-(trifluoromethoxy)phenylhydrazone (FCCP) titration. Some potential applications of this oxygen sensor system include evaluating effects of metabolic therapies on oocyte bioenergetics, and monitoring mitochondrial function throughout oocyte maturation and blastocyst development to predict embryo viability to compliment assisted reproductive technologies.

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“…The use of oxygen sensors for in-vitro monitoring of cell activities and for determining the effect of drugs on cell metabolism has been demonstrated using optical techniques such as florescence quenching by measuring the change in oxygen concentration near the cell [12,13]. This method has drawbacks such as temperature sensitivity, and it requires a long time to reach stability [13,14].…”

Section: Introductionmentioning

confidence: 99%

Monitoring of Oocyte/Embryo Respiration Using Electrochemical-Based Oxygen Sensors

Obeidat¹,

Evans²,

Tedjo³

et al. 2017

Preprint

0

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Current commercially available instruments for monitoring mitochondrial respiration are incapable of single cell measurements. Therefore, we developed a three-electrode, Clark-type biosensor suitable for mitochondrial respirometry in single oocytes and embryos. The biosensor was embedded in a PMMA (polymethyl methacrylate) micro-chamber to allow investigation of single oocytes/embryos immersed in up to 100 µL of respiration buffer. The micro-chamber was completely sealed to avoid oxygen exchange between the inside of the chamber and the atmosphere, while being maintained at a temperature of 38.5 ˚C to preserve cell viability. Using amperometry, the oxygen consumption of cells inside the micro-chamber was measured as a change in output current and converted to femto-mol (fmol) oxygen consumed per second based on calibrations with known buffer oxygen concentrations. The sensor measured basal cell respiration supported by endogenous substrates, respiration associated with proton leak induced by inhibition of the adenosine triphosphate (ATP) synthase (complex V) with oligomycin, and the maximal noncoupled respiratory capacity revealed by Carbonyl cyanide-4-(trifluoromethoxy)phenylhydrazone (FCCP) titration. Some potential applications of this oxygen sensor system include evaluating effects of metabolic therapies on oocyte bioenergetics, and monitoring mitochondrial function throughout oocyte maturation and blastocyst development to predict embryo viability to compliment assisted reproductive technologies.

show abstract