Developments in cell biology for quantitative immunoelectron microscopy based on thin sections: a review.
Developments in cell biology for quantitative immunoelectron microscopy based on thin sections: a review.
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DOI：
10.1007/s00418-008-0451-6
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发表时间：
2008-08
影响因子：
2.3
通讯作者：
Lucocq, John M.
中科院分区：
文献类型：
--
作者：
Mayhew, Terry M.;Lucocq, John M.
关键词：
定量免疫电子显微镜使用荧光切片和金颗粒标记来确定分子在细胞隔室中的分布。在这里，我们回顾了一系列新方法，用于比较一个研究组（方法1）中不同隔室之间以及两个或多个组（方法2）中相同隔室之间的标记分布。无偏地选择样本，然后通过适当的统计程序估计和比较观察到的和预期的金颗粒分布。该方法可用于分析分布在体积占据（细胞器）和表面占据（膜）室之间的金标记，但在方法1中，膜必须作为细胞器处理。使用方法1，将金计数与室大小的体视学估计相结合以确定标记密度（LD）。对于体积占用者，LD可以简单地表示为每个测试点的金数，对于表面占用者，表示为每个测试线交叉点的金数。通过将金颗粒随机分配到隔室并将观察到的/预期的计数表示为相对标记指数（RLI）来生成预期分布。优先标记的车厢确定从他们的RLI值和卡方分析观察到的和预期的分布。对于方法2，通过列联表和卡方分析简单地比较各组间分布在隔室之间的原始金颗粒计数。这确定了负责组分布之间的差异的主要隔间。最后，我们讨论了标记效率（每个目标分子的金颗粒的数量），并描述了如何可以估计体积或表面占位相结合的体视学数据与生化测定。
Quantitative immunoelectron microscopy uses ultrathin sections and gold particle labelling to determine distributions of molecules across cell compartments. Here, we review a portfolio of new methods for comparing labelling distributions between different compartments in one study group (method 1) and between the same compartments in two or more groups (method 2). Specimen samples are selected unbiasedly and then observed and expected distributions of gold particles are estimated and compared by appropriate statistical procedures. The methods can be used to analyse gold label distributed between volume-occupying (organelle) and surface-occupying (membrane) compartments, but in method 1, membranes must be treated as organelles. With method 1, gold counts are combined with stereological estimators of compartment size to determine labelling density (LD). For volume-occupiers, LD can be expressed simply as golds per test point and, for surface-occupiers, as golds per test line intersection. Expected distributions are generated by randomly assigning gold particles to compartments and expressing observed/expected counts as a relative labelling index (RLI). Preferentially-labelled compartments are identified from their RLI values and by Chi-squared analysis of observed and expected distributions. For method 2, the raw gold particle counts distributed between compartments are simply compared across groups by contingency table and Chi-squared analysis. This identifies the main compartments responsible for the differences between group distributions. Finally, we discuss labelling efficiency (the number of gold particles per target molecule) and describe how it can be estimated for volume- or surface-occupiers by combining stereological data with biochemical determinations.
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影响因子：
2
作者：
Gundersen, HJG;Jensen, EBV;Nielsen, J
通讯作者：
Nielsen, J
影响因子：
56.9
作者：
Bendayan, M
通讯作者：
Bendayan, M
影响因子：
1.6
作者：
Borjigin, Gerelt;Yamamoto, Shuhei;Suzuki, Atsushi
通讯作者：
Suzuki, Atsushi
DOI：
10.1111/j.1365-2362.2006.01732.x
发表时间：
2006-12-01
影响因子：
5.5
作者：
Li, A. C. Y.;Warley, A.;Powell, J. J.
通讯作者：
Powell, J. J.
影响因子：
4.3
作者：
Lopes, Marta S.;Cortadellas, Nuria;Araus, Jose L.
通讯作者：
Araus, Jose L.