Division of Plastic and Reconstructive Surgery, Department of Surgery, Washington University School of Medicine in Saint Louis, St Louis, MO, 63110, USA.
Department of Plastic and Reconstructive Surgery, The Ohio State University, Wexner Medical Center, 915 Olentangy River Rd, Suite 2100, Columbus, OH, 43212, USA.
J Neurosci Methods. 2020 Apr 15;336:108635. doi: 10.1016/j.jneumeth.2020.108635. Epub 2020 Feb 15.
Stereology and histomorphometry are widely used by investigators to quantify nerve characteristics in normal and pathological states, including nerve injury and regeneration. While these methods of analysis are complementary, no study to date has systematically compared both approaches in peripheral nerve. This study investigated the reliability of design-based stereology versus semi-automated binary imaging histomorphometry for assessing healthy peripheral nerve characteristics.
Stereological analysis was compared to histomorphometry with binary image analysis on uninjured sciatic nerves to determine nerve fiber number, nerve area, neural density, and fiber distribution.
Sciatic nerves were harvested from 6 male Lewis rats, aged 8-12 weeks for comprehensive analysis of 6 nerve specimens. From each animal, sciatic nerve specimens were fixed, stained, and sectioned for analysis by light and electron microscopy. Both histomorphometry and stereological peripheral nerve analyses were performed on all specimens by two blinded and independent investigators who quantified nerve fiber count, fiber width, density, and related distribution parameters.
Histomorphometry and stereological analysis provided similar outcomes in nerve fiber number and total nerve area. However, the light microscopy, but not electron microscopy, stereological analysis yielded higher nerve fiber area compared to histomorphometry or manual measurement.
Both methods measure similar fiber number and overall nerve fiber area; however, stereology with light microscopy quantified higher fiber area. Histomorphometry optimizes throughput and comprehensive analysis but requires user thresholding.
体视学和组织形态计量学被广泛用于研究人员在正常和病理状态下定量神经特征，包括神经损伤和再生。虽然这些分析方法是互补的，但迄今为止尚无研究系统比较外周神经中的这两种方法。本研究旨在调查基于设计的体视学与半自动二值图像组织形态计量学在评估健康周围神经特征方面的可靠性。
在未受伤的坐骨神经上比较体视学与二值图像分析的组织形态计量学，以确定神经纤维数量、神经面积、神经密度和纤维分布。
从 6 只 8-12 周龄雄性 Lewis 大鼠中采集坐骨神经，对 6 个神经标本进行全面分析。从每只动物中取出坐骨神经标本，进行固定、染色和切片，用于光镜和电镜分析。两位盲法独立的研究者对所有标本进行了组织形态计量学和体视学周围神经分析，对神经纤维计数、纤维宽度、密度和相关分布参数进行了定量。
组织形态计量学和体视学分析在外周神经纤维数量和总神经面积方面提供了相似的结果。然而，与组织形态计量学或手动测量相比，光镜体视学分析得出的神经纤维面积高于组织形态计量学。
两种方法测量的纤维数量和总神经纤维面积相似；然而，光镜体视学定量了更高的纤维面积。组织形态计量学优化了吞吐量和全面分析，但需要用户进行阈值处理。
