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首页> 外文期刊>Plant Disease >An Improved Technique for Sorting Developmental Stages and Assessing Egg Viability of Globodera pallida using High-Throughput Complex Object Parametric Analyzer and Sorter
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An Improved Technique for Sorting Developmental Stages and Assessing Egg Viability of Globodera pallida using High-Throughput Complex Object Parametric Analyzer and Sorter

机译：用高通量复杂对象参数分析仪分析发展阶段的改进技术，评估Gloomodera Pallida的鸡蛋活力

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摘要

The Complex Object Parametric Analyzer and Sorter (COPAS) is a large particle flow cytometer designed for analyzing, sorting, and dispensing objects of varying sizes. We explored the potential of using this instrument to analyze and sort various developmental stages and egg viability of Globodera pallida. Cysts were successfully examined and sorted from debris by optimizing side-scatter and red-fluorescence parameters on the COPAS. We were able to separate eggs and second-stage juveniles from samples of mixed population using extinction and time of flight. Separation of live and dead eggs was examined following staining eggs with SYTOX Green and application of time of flight and green peak height. Data were compared with a commonly used viability assay by which eggs were stained with Meldola's Blue and examined by a microscope. COPAS proved to be effective in assessing viability by detecting two separate gates: live eggs having green fluorescence peaks &190 and dead eggs with the peaks &190. The application of COPAS in combination with SYTOX Green detected a greater number of live eggs than the Meldola's assay, suggesting that SYTOX Green provided an overestimate of live eggs. COPAS noticeably increased the accuracy and reduced the time required for screening and analyzing nematode populations.
机译：复杂对象参数分析仪和分类器（COPAS）是一种大的颗粒流动细胞仪，专门用于分析，分类和分配不同尺寸的物体。我们探讨了使用这种仪器的潜力来分析和分析Globodera Pallida的各种发育阶段和鸡蛋活力。通过优化COPAS上的侧散散和红荧光参数，成功检查并从碎片中检查并分类。我们能够使用灭绝和飞行时间将鸡蛋和二阶青少年分离鸡蛋和二阶青少年。在染色鸡蛋与Sytox绿色和飞行时间和绿色峰值高度的应用，检查了活生和死蛋的分离。将数据与常用的活力测定进行比较，通过将蛋染成米尔多拉的蓝色并通过显微镜检查。 Copas证明通过检测两个单独的门来评估活力：生物彩蛋，具有绿色荧光峰＆amp; LT; 190和峰值的死蛋; GT; 190。 COPAS与Sytox Green的应用相结合地检测到比Meldola的测定更大量的活蛋，表明Sytox Green提供了过度的活蛋。 COPAS明显增加了准确性并减少了筛选和分析线虫种群所需的时间。

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