Unbiased estimation of cell number using the automatic optical fractionator.
Unbiased estimation of cell number using the automatic optical fractionator.
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DOI：
10.1016/j.jchemneu.2016.12.002
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发表时间：
2017-03
影响因子：
2.8
通讯作者：
Morgan D
中科院分区：
文献类型：
--
作者：
Mouton PR;Phoulady HA;Goldgof D;Hall LO;Gordon M;Morgan D
一种新的体视学方法，自动光学分馏器，提出了获得无偏和有效的估计组织切片中的细胞数量。与现有的分割算法和普通的免疫染色方法结合使用，可以从由三维组织体积（解剖器堆栈）构建的扩展景深图像中获得细胞数量的自动估计。自动光学分馏仪更准确，100%客观，比手动光学分馏仪快8到10倍。提供了自动分级器的一个实例，用于神经变性的遗传修饰小鼠模型的新皮质中免疫染色神经元的计数。证据是经常被忽视的先决条件，精确计数的光学分馏器需要一个薄的焦平面产生的高光学分辨率透镜。
A novel stereology approach, the automatic optical fractionator, is presented for obtaining unbiased and efficient estimates of the number of cells in tissue sections. Used in combination with existing segmentation algorithms and ordinary immunostaining methods, automatic estimates of cell number are obtainable from extended depth of field images built from three-dimensional volumes of tissue (disector stacks). The automatic optical fractionator is more accurate, 100% objective and 8 to 10 times faster than the manual optical fractionator. An example of the automatic fractionator is provided for counts of immunostained neurons in neocortex of a genetically modified mouse model of neurodegeneration. Evidence is presented for the often overlooked prerequisite that accurate counting by the optical fractionator requires a thin focal plane generated by a high optical resolution lens.