公开(公告)号：EP2916131A1

公开(公告)日：2015-09-09

申请号：EP14157928.4

申请日：2014-03-05

申请人： Amphasys AG

发明人： Di Berardino, Marco ,  Schade-Kampmann, Grit

IPC分类号： G01N33/04 ,  A01J5/013 ,  G01N15/12

CPC分类号： G01N33/48735 ,  A01J5/0132 ,  A01J5/0133 ,  G01N15/1031 ,  G01N15/1056 ,  G01N33/04 ,  G01N33/48785

摘要： The present invention refers to a method for the discrimination of cells from other particles, as well as of different types of cells in raw milk samples by impedance microflow cytometry. More precisely, it describes a method of analysing the quality of raw milk in terms of its bacterial and somatic cell content without the need to pre-treat the milk sample, such that the analysis can take place directly at the production site. One advantage of the invention is discriminating and counting somatic cells from milk particles (consisting mainly of lipid vesicles) by highfrequency impedance analysis directly performed on untreated raw milk. Another advantage of this invention is that the method allows diagnosing the status of a mastitis infection directly after milking according to the analysis of the somatic cell count. Another advantage of this invention is to allow a fast method for determining the bacterial cell count in raw milk directly after milking. In addition, viability of both somatic and bacterial cells can be determined without the need of any cell label. A further advantage is, that the analysis can be obtained in realtime, directly after the raw milk has passed the micro channel of the microfluidic device.

摘要翻译： 本发明涉及通过阻抗微流式细胞术鉴别生奶样品中其它颗粒以及不同类型细胞的细胞的方法。 更准确地说，它描述了一种以细菌和体细胞含量为基础来分析生奶的质量的方法，而不需要预处理牛奶样品，使得分析可以直接在生产现场进行。 本发明的一个优点是通过直接在未经处理的生牛奶上进行的高频阻抗分析来区分并计数来自乳颗粒（主要由脂质囊泡组成）的体细胞。 本发明的另一个优点是该方法允许根据体细胞计数的分析直接在挤奶之后诊断乳腺炎感染的状态。 本发明的另一个优点是允许在挤奶后直接测定生奶中的细菌细胞计数的快速方法。 此外，可以确定体细胞和细菌细胞的活力，而不需要任何细胞标记。 另一个优点是，可以在生奶通过微流体装置的微通道之后直接获得实时分析。

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。

公开(公告)号：EP2916131B1

公开(公告)日：2018-01-17

申请号：EP14157928.4

申请日：2014-03-05

申请人： Amphasys AG

发明人： Di Berardino, Marco ,  Schade-Kampmann, Grit

IPC分类号： G01N33/04 ,  A01J5/013 ,  G01N15/12

CPC分类号： G01N33/48735 ,  A01J5/0132 ,  A01J5/0133 ,  G01N15/1031 ,  G01N15/1056 ,  G01N33/04 ,  G01N33/48785

摘要： The present invention refers to a method for the discrimination of cells from other particles, as well as of different types of cells in raw milk samples by impedance microflow cytometry. More precisely, it describes a method of analysing the quality of raw milk in terms of its bacterial and somatic cell content without the need to pre-treat the milk sample, such that the analysis can take place directly at the production site. One advantage of the invention is discriminating and counting somatic cells from milk particles (consisting mainly of lipid vesicles) by highfrequency impedance analysis directly performed on untreated raw milk. Another advantage of this invention is that the method allows diagnosing the status of a mastitis infection directly after milking according to the analysis of the somatic cell count. Another advantage of this invention is to allow a fast method for determining the bacterial cell count in raw milk directly after milking. In addition, viability of both somatic and bacterial cells can be determined without the need of any cell label. A further advantage is, that the analysis can be obtained in realtime, directly after the raw milk has passed the micro channel of the microfluidic device.