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公司名称: Carl Roth
产品编号: 4256.2
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Quantitative Image Analysis of Membrane Microdomains Labelled by Fluorescently Tagged Proteins in Arabidopsis thaliana and Nicotiana benthamiana
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2015-06-05
[Abstract]  We have recently characterized co-existing membrane microdomains that are labeled by different proteins in living plant cells (Jarsch et al., 2014). For this approach we first created a digital fingerprint for each of the twenty marker proteins using quantitative image analysis. Here we recorded parameters such as domain size, density and shape based on image segmentation. We found highly reproducible patterns of any of the proteins over a large number of biological replicates. Furthermore we exclusively acquired images from lowly expressing cells and chose our imaging conditions in ... [摘要]  我们最近描述了在活植物细胞中由不同蛋白质标记的共存膜微结构域(Jarsch等人,2014)。对于这种方法,我们首先使用定量图像分析为二十个标记蛋白质中的每一个创建数字指纹。这里我们基于图像分割记录诸如域大小,密度和形状的参数。我们发现任何蛋白质的高度可重复的模式在大量的生物复制。此外,我们专门从低表达细胞中获取图像,并选择我们的成像条件以产生没有像素饱和的图像。
该方案详细描述了用于分析成员的定位差异的方法的拟南芥(Arabidopsis thaliana)和本塞姆氏烟草(Nicotiana benthamiana)的膜微结构域中的remorin蛋白家族(Jarsch等人,2014)。蛋白质单独地或成对地表达为相应植物中的荧光团融合体。使用标准共焦激光扫描显微镜(CLSM)进行图像采集,并使用ImageJ进行图像分析。

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