Burke Mark, Zangenehpour Shahin, Mouton Peter R, Ptito Maurice
Department of Physiology, University of Montreal.
J Vis Exp. 2009 May 14(27):1262. doi: 10.3791/1262.
The non-human primate is an important translational species for understanding the normal function and disease processes of the human brain. Unbiased stereology, the method accepted as state-of-the-art for quantification of biological objects in tissue sections, generates reliable structural data for biological features in the mammalian brain. The key components of the approach are unbiased (systematic-random) sampling of anatomically defined structures (reference spaces), combined with quantification of cell numbers and size, fiber and capillary lengths, surface areas, regional volumes and spatial distributions of biological objects within the reference space. Among the advantages of these stereological approaches over previous methods is the avoidance of all known sources of systematic (non-random) error arising from faulty assumptions and non-verifiable models. This study documents a biological application of computerized stereology to estimate the total neuronal population in the frontal cortex of the vervet monkey brain (Chlorocebus aethiops sabeus), with assistance from two commercially available stereology programs, BioQuant Life Sciences and Stereologer (Figure 1). In addition to contrast and comparison of results from both the BioQuant and Stereologer systems, this study provides a detailed protocol for the Stereologer system.
非人灵长类动物是理解人类大脑正常功能和疾病过程的重要转化研究物种。无偏倚立体学是组织切片中生物物体定量的公认先进方法,可为哺乳动物大脑的生物学特征生成可靠的结构数据。该方法的关键组成部分是对解剖学定义结构(参考空间)进行无偏倚(系统随机)采样,并结合对细胞数量和大小、纤维和毛细血管长度、表面积、区域体积以及参考空间内生物物体的空间分布进行定量。与先前方法相比,这些立体学方法的优势之一在于避免了因错误假设和不可验证模型而产生的所有已知系统(非随机)误差来源。本研究记录了在两款市售立体学程序BioQuant Life Sciences和Stereologer的辅助下,计算机化立体学在估计绿猴(Chlorocebus aethiops sabeus)大脑额叶皮质神经元总数方面的生物学应用(图1)。除了对比和比较BioQuant和Stereologer系统的结果外,本研究还提供了Stereologer系统的详细方案。