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

Xia, Xiufang; Wan, Yinglang; Wang, Wenjun; Yin, Guojun; McLamore, Eric S.; Lu, Xianju

doi:10.1038/srep03057

Cited by 25 publications

(20 citation statements)

References 19 publications

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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%

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%

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

et al. 2020

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“…Scientists from China have developed a new microprototype technique (a highly sensitive and selective method for measuring the concentration and flux of oxygen on the cell surface) [11] for a quick and non-invasive measurement of seed viability by measuring the oxygen supply to rT, relative temperature; (Left y axes and gray time windows) Rate with which the viability of an unknown seed was successfully predicted. (Middle y axes and black lines) significance levels with which the mean values of two seed types differed over time.…”

Section: Experimental Partmentioning

confidence: 99%

Trends of the development of methods for determining the viability of crop seeds

Moskovskiy¹,

Khamuev²,

Lepeshkin³

2019

Proceedings of the International Scientific and Practical Conference “Digital Agriculture - Development Strategy” (ISPC 2019)

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Scientific research to determine the viability of the system of cleaning and sorting of seeds. An analysis of the current state of determining seed viability has shown that, despite many studies, the main task of promising research projects is the development of a high-precision X-ray laser, a device that uses stimulated radiation to generate or amplify electromagnetic radiation in the near-X-ray or ultraviolet region of the spectrum, that is, as a rule, about several tens of nanometers (nm) wavelength. The main results of the use of a set of technical tools based on Raman spectroscopy laser is the determination of seed viability in a short period of time and obtaining information about the ability of seeds to germinate. Substantiate the need to use laser Raman spectroscopy to identify a complex of biomarkers of crop seeds as signs of their viability, as well as for computer (intelligent) electron-optical cleaning and sorting of seeds. The biological properties of seeds must be considered when developing the process of sowing seed. Among the basic biological properties of seeds include viability. Existing traditional technical means for determining the viability of crop seeds do not fully cope with the task.

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“…Presently predominant methods for monitoring viability in seed banks are germination test, 2,3,5-triphenyltetrazolium hydrochloride dyeing, and electric conductivity measurement [ 13 ]. But germination is the overwhelmingly widespread one owing the following advantages (1) precision (successful transition to a seedling means that the seed is viable); (2) simplicity (easy to operate) [ 14 ].…”

Section: Introductionmentioning

confidence: 99%

“…But germination is the overwhelmingly widespread one owing the following advantages (1) precision (successful transition to a seedling means that the seed is viable); (2) simplicity (easy to operate) [ 14 ]. Despite its usefulness, this method has drawbacks: post-germination seeds cannot be stored as seeds anymore, and the duration may be too long for early decision on whether an accession should be stored as usual, primed to enhance germination or regenerated [ 13 ]. Masses of up-to-date research in this decade focus on the two problems, with the advantage described as either “non-invasive/non-destructive” (N) or “real-time/rapid” (R), featured with sensing (S) apparatus (NRS method, NRSs) [ 14 – 18 ].…”

Section: Introductionmentioning

confidence: 99%

Dehiscence method: a seed-saving, quick and simple viability assessment in rice

Lin

Wang

et al. 2018

Plant Methods

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BackgroundSeed viability monitoring is very important in ex situ germplasm preservation to detect germplasm deterioration. This requires seed-, time- and labor- saving methods with high precision to assess seed germination as viability. Although the current non-invasive, rapid, sensing methods (NRSs) are time- and labor-saving, they lack the precision and simplicity which are the virtues of traditional germination. Moreover, they consume a considerable amount of seeds to adjust sensed signals to germination percentage, which disregards the seed-saving objective. This becomes particularly severe for rare or endangered species whose seeds are already scarce. Here we propose a new method that is precise, low-invasive, simple, and quick, which involves analyzing the pattern of dehiscence (seed coat rupture), followed by embryonic protrusion.ResultsDehiscence proved simple to identify. After the trial of 20 treatments from 3 rice varieties, we recognized that dehiscence percentage at the 48th hour of germination (D(48)) correlates significantly with germination rate for tested seed lots. In addition, we found that the final germination percentage corresponded to D(48) plus 5. More than 70% of the seeds survived post-dehiscence desiccation for storage. Hydrogen peroxide (1 mM) as the solution for imbibition could further improve the survival. The method also worked quicker than tetrazolium which is honored as a fast, traditional method, in detecting less vigorous but viable seeds.ConclusionWe demonstrated the comprehensive virtues of dehiscence method in assessing rice seed: it is more precise and easier to use than NRSs and is faster and more seed-saving than traditional methods. We anticipate modifications including artificial intelligence to extend our method to increasingly diverse circumstances and species.Electronic supplementary materialThe online version of this article (10.1186/s13007-018-0334-3) contains supplementary material, which is available to authorized users.

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

Cited by 25 publications

References 19 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

Trends of the development of methods for determining the viability of crop seeds

Dehiscence method: a seed-saving, quick and simple viability assessment in rice

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