Ballou Edmund W, Smith W Bryan, Anelli Roberta, Heckman C J
Department of Physiology M211, Northwestern University Feinberg School of Medicine, Chicago, IL 60611, USA.
J Neurosci Methods. 2006 Sep 30;156(1-2):257-66. doi: 10.1016/j.jneumeth.2006.03.014. Epub 2006 May 9.
Neurons perform much of their integrative work in the dendritic tree, and spinal motoneurons have the largest tree of any cell. Electrical excitability is strongly influenced by dendrite membrane properties, which are difficult to measure directly. We describe a method to measure the distribution of ion channel membrane densities along dendritic trajectories. The method combines standard immunohistochemistry with reconstruction procedures for both large-scale and small-scale optical microscopy. Software written for Matlab then extracts the colocalization of the target ion channel with the target dye injected cell, and calculates the relative channel density per square micron of cell surface area, as a function of distance from the cell body. The technique can be used to quantify the localization and distribution of any immunoreactive moiety, and the software provides a flexible vehicle for sensitivity analysis, to validate heuristics for selecting thresholds.
神经元在树突中进行大量的整合工作，而脊髓运动神经元拥有所有细胞中最大的树突。电兴奋性受树突膜特性的强烈影响，而树突膜特性很难直接测量。我们描述了一种测量沿树突轨迹的离子通道膜密度分布的方法。该方法将标准免疫组织化学与用于大规模和小规模光学显微镜的重建程序相结合。然后，为Matlab编写的软件提取目标离子通道与注入目标染料的细胞的共定位，并计算每平方微米细胞表面积的相对通道密度，作为距细胞体距离的函数。该技术可用于量化任何免疫反应性部分的定位和分布，并且该软件为敏感性分析提供了一个灵活的工具，以验证选择阈值的启发式方法。
