公开(公告)号：EP2915423A1

公开(公告)日：2015-09-09

申请号：EP14158150.4

申请日：2014-03-06

申请人： Amphasys AG

发明人： Di Berardino, Marco ,  Schade-Kampmann, Grit ,  Kok, Kim ,  Heimann, Iris

IPC分类号： A01H1/02 ,  A01H1/04

CPC分类号： G01N27/20 ,  A01H1/025 ,  A01H1/04 ,  G01N33/4833

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.

摘要翻译： 用于确定花粉种群花粉活力和/或成熟等级的方法包括：从花中机械去除花粉; 将花粉粒重新悬浮在导电缓冲液中，以保持植物细胞保持活力; 将花粉悬浮液通过适合于适于进行阻抗流式细胞术（IFC）的微流体装置的孔径的合适的过滤器; 将经过滤的悬浮液通过微流体装置并测量花粉粒的阻抗; 将每个花粉的测量阻抗值与振幅值及其相关相位角以及测量频率进行存储; 计算过滤悬浮液中的花粉粒; 以及通过设定适当的门并确定所分析的花粉种群的存活率或成熟等级，根据幅度和/或相位值来区分花粉粒。 本发明的一个优点是，植物细胞不必对生存力分析进行染色，该方法是非侵入性的，并且可以在短时间内实时和现场分析大量细胞。 可以跟踪植物细胞的发育过程，适用于所有植物物种。 特别地，该方法允许独立于位置和世界各地的标准测量。