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Method for the determination of pollen viability and maturation grade of a pollen population

机译:测定花粉群的花粉活力和成熟等级的方法

摘要

The method for the determination of pollen viability and/or maturation grade of a pollen population, comprises: mechanically removing pollen grains from flowers; re-suspending pollen grains in an electrically conductive buffer for keeping the plant cells to remain viable; passing the pollen suspension through an appropriate filter with a pore size suitable for a microfluidic device being adapted to perform impedance flow cytometry (IFC); passing the filtered suspension through the microfluidic device and measuring an impedance of the pollen grains; storing the measured impedance values of each pollen with the amplitude value and its correlated phase angle as well as the measurement frequencies; counting the pollen grains in the filtered suspension; and discriminating the pollen grains according to the amplitude and/or phase values by setting an appropriate gate and determining the viability ratio or maturation grade of the analysed pollen population. An advantage of the present invention is that a plant cell does not have to be stained for viability analysis, the method is non-invasive and a high number of cells can be analysed in a short time frame in real-time and on-site. It is possible to follow developmental processes of plant cells, and that it is applicable to all plant species. In particular, the method allows a standardized measurement independent of the location and all over the world.
机译:用于测定花粉群的花粉活力和/或成熟等级的方法包括:机械地从鲜花中除去花粉粒;重新悬浮在导电缓冲液中的花粉颗粒,以保持植物细胞保持可行;通过花粉悬浮液通过适当的过滤器,其具有适于进行微流体装置的孔径,适于进行阻抗流式细胞术(IFC);通过微流体装置通过过滤的悬浮液并测量花粉颗粒的阻抗;将每个花粉的测量阻抗值与幅度值及其相关的相角以及测量频率存储;计算过滤悬浮液中的花粉颗粒;通过设定适当的浇口并确定分析的花粉群的可存活率或成熟级别来根据幅度和/或相值来区分花粉颗粒。本发明的一个优点是植物细胞不必染色用于活性分析,该方法是非侵入性的,并且可以在实时和现场在短时间帧中分析大量细胞。可以追随植物细胞的发育过程,并且它适用于所有植物物种。特别是,该方法允许标准化测量与世界各地无关。

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