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首页> 外文期刊>Journal of Neuroscience Methods >Calibration of the stereological estimation of the number of myelinated axons in the rat sciatic nerve: a multicenter study.
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Calibration of the stereological estimation of the number of myelinated axons in the rat sciatic nerve: a multicenter study.

机译：大鼠坐骨神经中髓鞘轴突数量的立体估计的校准：一项多中心研究。

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摘要

Several sources of variability can affect stereological estimates. Here we measured the impact of potential sources of variability on numerical stereological estimates of myelinated axons in the adult rat sciatic nerve. Besides biological variation, parameters tested included two variations of stereological methods (unbiased counting frame versus 2D-disector), two sampling schemes (few large versus frequent small sampling boxes), and workstations with varying degrees of sophistication. All estimates were validated against exhaustive counts of the same nerve cross sections to obtain calibrated true numbers of myelinated axons (gold standard). In addition, we quantified errors in particle identification by comparing light microscopic and electron microscopic images of selected consecutive sections. Biological variation was 15.6%. There was no significant difference between the two stereological approaches or workstations used, but sampling schemes with few large samples yielded larger differences (20.7+/-3.7% SEM) of estimates from true values, while frequent small samples showed significantly smaller differences (12.7+/-1.9% SEM). Particle identification was accurate in 94% of cases (range: 89-98%). The most common identification error was due to profiles of Schwann cell nuclei mimicking profiles of small myelinated nerve fibers. We recommend sampling frequent small rather than few large areas, and conclude that workstations with basic stereological equipment are sufficient to obtain accurate estimates. Electron microscopic verification showed that particle misidentification had a surprisingly variable and large impact of up to 11%, corresponding to 2/3 of the biological variation (15.6%). Thus, errors in particle identification require further attention, and we provide a simple nerve fiber recognition test to assist investigators with self-testing and training.
机译：可变性的几种来源可能会影响立体估计。在这里，我们测量了成年大鼠坐骨神经中髓鞘轴突的数字立体估计的潜在变异源的影响。除生物学差异外，所测试的参数还包括两种立体方法的变化（无偏计数框架与2D解剖器），两种采样方案（很少有大型采样箱与经常使用的小型采样箱）以及具有不同复杂程度的工作站。所有估计都针对相同神经横断面的穷举计数进行验证，以获得经过校准的有髓神经轴突的真实数量（金标准）。此外，我们通过比较选定连续部分的光学显微镜和电子显微镜图像来量化颗粒识别中的误差。生物变异为15.6％。所使用的两种立体学方法或工作站之间没有显着差异，但是使用少量大样本的采样方案与真实值相比，估计值产生较大的差异（20.7 +/- 3.7％SEM），而频繁的小样本显示出明显较小的差异（12.7+ /-1.9% SEM）。在94％的病例中，颗粒物识别准确（范围：89-98％）。最常见的识别错误是由于雪旺细胞核的轮廓模仿了小髓神经纤维的轮廓。我们建议对小区域（而不是大区域）进行频繁采样，并得出结论，配备基本立体音响设备的工作站足以获得准确的估计值。电子显微镜验证表明，颗粒物的误识别具有令人惊讶的可变性，且影响高达11％，相当于生物学变化的2/3（15.6％）。因此，颗粒识别中的错误需要进一步关注，我们提供了一种简单的神经纤维识别测试，以协助研究人员进行自我测试和培训。

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