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首页> 外文期刊>Histochemistry and cell biology >Applications of an efficient method for comparing immunogold labelling patterns in the same sets of compartments in different groups of cells.
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Applications of an efficient method for comparing immunogold labelling patterns in the same sets of compartments in different groups of cells.

机译：一种有效方法的应用，用于比较不同细胞组中同一组隔室中的免疫金标记模式。

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摘要

Quantitative immunoelectron microscopy often involves determining the distributions of gold label in different intracellular compartments and then drawing comparisons between compartments in the same sample of cells or between experimental groups of cells. In the case of within-group comparisons, recent developments in the estimation of relative labelling index and labelling density make it possible to test whether or not particular compartments are preferentially labelled. These methods are ideally suited to analysing gold label restricted to volume (organelle) or surface (membrane) compartments but may be modified to analyse label localised in mixtures of both. Here, a simple and efficient approach to drawing between-group comparisons for label associated with organelles and/or membranes is presented. The method relies on multistage random sampling of specimens (via blocks and microscopic fields) followed by simply counting gold particles associated with different compartments. The distributions of raw gold counts in different groups are then compared by contingency table analysis with statistical degrees of freedom for chi-squared values being determined by the number of compartments and the number of experimental groups of cells. Compartmental chi-squared values making substantial contributions to the total chi-squared values then identify where the main between-group differences reside. The method requires no information about compartment size (for example, organelle profile area or membrane trace length) and does not even depend critically on standardising between-group magnification. Its application is illustrated using datasets from immunolabelling studies designed to localise the KDEL receptor, phosphatidyl-inositol 4,5-bisphosphate, GLUT4 and rab4 at the electron microscopic level.
机译：定量免疫电子显微镜检查通常涉及确定金标记在不同细胞内区室中的分布，然后在同一细胞样品中的区室之间或实验组细胞之间进行比较。在组内比较的情况下，在估计相对标记指数和标记密度方面的最新进展使得有可能测试特定的区室是否被优先标记。这些方法非常适合分析限制在体积（细胞器）或表面（膜）隔室中的金标，但可以进行修改以分析定位在两者混合物中的金标。在此，提出了一种简单有效的方法来绘制与细胞器和/或膜相关的标记的组间比较。该方法依赖于样品的多阶段随机采样（通过块和微观区域），然后简单地计数与不同隔室相关的金颗粒。然后，通过列联表分析，比较不同组中原金计数的分布，并根据隔室数和实验组细胞数确定卡方值的统计自由度。间隔卡方值对总卡方值有重大贡献，然后确定组间主要差异所在的位置。该方法不需要有关隔室大小的信息（例如，细胞器轮廓区域或膜迹线长度），甚至根本不依赖于标准化组间放大倍数。使用免疫标记研究的数据集说明了其应用，该数据集旨在在电子显微镜水平上定位KDEL受体，磷脂酰肌醇4,5-双磷酸酯，GLUT4和rab4。

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