Non-invasive measurement of real-time oxygen flux in plant systems with a self-referencing optrode

Wan, Yinglang; McLamore, Eric S.; Fan, Lusheng; Hao, Huaiqing; Porterfield, D. Marshall; Zhang, Zengkai; Wang, Wenjun; Xu, Yan; Lin, Jingkai

doi:10.1038/protex.2011.266

Cited by 4 publications

(5 citation statements)

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“…Technological advancements in optical fibers, light sources (including lightemitting diodes (LEDs)), photodiodes, cameras, motion control and software decreased the cost and improved the sensor system, progressing it to the modern, herein-described, state 44 . These developments broadened the use of optrodes to measure O 2 fluxes, with an emphasis on plant physiology, for which various research and technical essays appeared 28,43,[45][46][47][48][49] . Our research studies, however, focus on animal physiology and are carried out using SMOT to measure the extracellular fluxes of dissolved O 2 (Fig.…”

Section: Development Of the Fluorescence Quenching-based Methodsmentioning

confidence: 99%

“…In our recordings, we optimized the sampling rules to 1-s wait time, 0.5-s sample time and 30-μm excursion, which gave a sampling interval of 11 s (~0.1 Hz). Acquisition frequencies around this value are common 46,48,49 and yield a good signal. Once optimized, these rules can be kept unchanged in all measurements.…”

Section: Box 3 | Sampling Rule Optimization For Data Acquisition In Asetmentioning

confidence: 98%

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Real-time physiological measurements of oxygen using a non-invasive self-referencing optical fiber microsensor

et al. 2020

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Reactive molecular oxygen (O 2) plays important roles in bioenergetics and metabolism and is implicated in biochemical pathways underlying angiogenesis, fertilization, wound healing and regeneration. Here we describe how to use the scanning micro-optrode technique (SMOT) to measure extracellular fluxes of dissolved O 2. The self-referencing O 2-specific micro-optrode (also termed micro-optode and optical fiber microsensor) is a tapered optical fiber with an O 2-sensitive fluorophore coated onto the tip. The O 2 concentration is quantified by fluorescence quenching of the fluorophore emission upon excitation with blue-green light. The micro-optrode presents high spatial and temporal resolutions with improved signal-to-noise ratio (in the picomole range). In this protocol, we provide step-by-step instructions for micro-optrode calibration, validation, example applications and data analysis. We describe how to use the technique for cells (Xenopus oocyte), tissues (Xenopus epithelium and rat cornea), organs (Xenopus gills and mouse skin) and appendages (Xenopus tail), and provide recommendations on how to adapt the approach to different model systems. The basic, user-friendly system presented here can be readily installed to reliably and accurately measure physiological O 2 fluxes in a wide spectrum of biological models and physiological responses. The full protocol can be performed in~4 h.

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Section: Development Of the Fluorescence Quenching-based Methodsmentioning

confidence: 99%

Section: Box 3 | Sampling Rule Optimization For Data Acquisition In Asetmentioning

confidence: 98%

Real-time physiological measurements of oxygen using a non-invasive self-referencing optical fiber microsensor

et al. 2020

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“…The MOT was calibrated in measuring solution with known oxygen concentrations (0 and 21%) by nitrogen and air purging at 20°C. The overall setup was designed according to previous literature717 to insure proper measurement of the concentration gradients from seeds. The equation of oxygen flux based on the changes of phase angel (φ) was: where J O2 is oxygen flux (pmol cm −2 s −1 ), D is molecular diffusion coefficient for oxygen in measuring solution (2.1 × 10 −5 cm 2 s −1 at 20°C), φ 1 is phase angle at near pole, φ 2 is phase angle at far pole, and m is linear slope of calibration plot within ambient conditions (0–21% O 2 ).…”

Section: Methodsmentioning

confidence: 99%

“…invented this technique (known generally as self referencingoptrodic sensing) and applied the tool to basic research for the measurement of oxygen fluxes in animal and plant systems13. Since that time, MOT has been utilized in a number of systems, including those for monitoring the effect of water quality on fish embryos by oxygen flux14, detecting chemical toxicity to bacterial biofilms15, determining plant root oxygen flux profiles7, screening auxin transport mutants16 and a protocol has even been established for the Arabidopsis root system17. The MOT is an ultrasensitive tool with high temporal and spatial resolution for detecting the physiological activity of live cells/tissues11.…”

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confidence: 99%

A real-time, non-invasive, micro-optrode technique for detecting seed viability by using oxygen influx

Xia

Wan

Wang³

et al. 2013

Sci Rep

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Quantifying seed viability is required for seed bank maintenance. The classical methods for detecting seed viability are time consuming and frequently cause seed damage and unwanted germination. We have established a novel micro-optrode technique (MOT) to measure seed viability in a quick and non-invasive manner by measuring the oxygen influxes of intact seeds, approximately 10 seconds to screen one seed. Here, we used soybean, wheat, and oilseed rape as models to test our method. After 3-hour imbibition, oxygen influxes were recorded in real-time with the total measurement taking less than 5 minutes. The results indicated a significantly positive correlation between oxygen influxes and viability in all 3 seed types. We also established a linear equation between oxygen influxes and seed viability for each seed type. For measurements, seeds were kept in the early imbibition stage without germination. Thus, MOT is a reliable, quick, and low-cost seed viability detecting technique.

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“…RS applies a microscope objective to focus the laser onto a specific sample and thus, is usually used to probe single cellular compositions (Huang et al, 2010 ). NMT is normally used to obtain molecular or ionic concentration and flux information of a small number of fixed cells or tissues (Zhang et al, 2009 ; Wan et al, 2011 ). Thus, RS and NMT techniques can help to track cellular compositional and physiological states easily.…”

Section: Introductionmentioning

confidence: 99%

Microalgae in Microwell Arrays Exhibit Differences with Those in Flasks: Evidence from Growth Rate, Cellular Carotenoid, and Oxygen Production

Ping

Yan

et al. 2018

Front. Plant Sci.

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Microalgae are cultivated in macro-scale reactors traditionally and the relevant knowledge is based on bulk analysis. Whether the knowledge and laws are true for cells under micro-cultivation is still unknown. To better understand microalgal physiology, micro-cultivation of microalgae, and unicellular tracking and analysis of its response in vivo is necessary. In the study, cellular responses of Chlorella vulgaris to micro-cultivation is studied, with cells in flasks as a control. Five different microwell depths ranging from 10 to 200 μm with a fixed diameter of 100 μm, and four diameter levels from 30 to 200 μm with a fixed depth 60 μm were investigated. Unicellular dynamics showed that cell number differences among various types of microwells with different initial cell numbers decreased as cultivation processed. Besides, the specific growth rate of C. vulgaris on microwell arrays was much higher than that in flasks and so cells on microwell arrsys can be much sensitive to pollutants. Thus, the interesting characteristics may be used in cell sensor applications to enhance sensitivity. The specific growth rate of C. vulgaris on microwell arrays decreased gradually as the microwell diameter increased from 30 to 200 μm while presented a unimodal trend as depth decreased from 200 to 10 μm. Furthermore, we used Raman Spectroscopy and Non-invasive Micro-test Technique to analyze cellular responses in microwells for the first time to track the changes in vivo. Results indicated that unicellular carotenoid content increased as microwells became larger and shallower. The flow rate of oxygen rose gradually as the depth increased from 10 to 100 μm, but then decreased rapidly as the depth deepened to 200 μm. In fact, it is a combined result of cell physiology and density. In summary, cells in microwells with the diameter/depth ratio ~1 owned the highest specific growth rates and oxygen flow rates. Simulations also suggested that better mass transfer occurred in microwells with higher diameter-to-depth ratios.

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Non-invasive measurement of real-time oxygen flux in plant systems with a self-referencing optrode

Cited by 4 publications

References 14 publications

Real-time physiological measurements of oxygen using a non-invasive self-referencing optical fiber microsensor

Real-time physiological measurements of oxygen using a non-invasive self-referencing optical fiber microsensor

A real-time, non-invasive, micro-optrode technique for detecting seed viability by using oxygen influx

Microalgae in Microwell Arrays Exhibit Differences with Those in Flasks: Evidence from Growth Rate, Cellular Carotenoid, and Oxygen Production

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