RESEARCH OF HISTOCHEMICAL STAINING FOR IDENTIFYING THE FUNCTION AND MORPHOLOGY OF FASCICLES IN THREE-DIMENSIONAL RECONSTRUCTION OF PERIPHERAL NERVES_Chinese Journal of Reparative and Reconstructive Surgery

Authors：

LUO Peng ¹ , ZHANG Yi ² , QI Jian ¹ , ZHONG Yingchun ³ , LIU Xiaolin. ¹

1Department of Orthopaedic Trauma and Microsurgery, 2Department of Plastic and Reconstructive Surgery, the First Affiliated Hospital of Sun Yat-sen University, Guangzhou Guangdong, 510080, P.R.China;;
3Faculty of Automation, Guangdong University of Technology.;

Corresponding author：

Keywords：

Peri pheral nerve; Three-dimensional reconstruction; Histochemical staining; Texture feature Image segmentation

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Objective To explore the histochemical staining for distinguishing and local izing nerve fibers and fascicles at histological level in three-dimensional reconstruction of peri pheral nerves. Methods The right median nerve was harvested from one fresh cadaver and embedded in OCT compound. The sample was serially horizontally sl iced with 6 μm thickness. All sections were stained with Karnovsky-Roots method (group A, n=30) firstly and then stained with toluidine blue (group B, =28) and Ponceau 2R (group C, n=21) in proper sequence. The results of each step were taken photos (× 100). After successfully stitching, the two-dimensional panorama images were compared, including texture feature, the number and aver gray level of area showing acetylchol inesterase (AchE) activity, and result of auto microscopic medical image segmentation. Results In groups A, B, and C, the number of AchE-positive area was (21.63 ± 4.06)× 102, (20.64 ± 3.51)× 102, and (20.54 ± 5.71)× 102, respectively, showing no significant difference among 3 groups (F=0.64, P=0.54); the mean gray level was (1.41 ± 0.06)× 102, (1.10 ± 0.05)× 102, and (1.14 ± 0.07)× 102, respectively, showing significant differences between group A and groups B and C (P lt; 0.001). In the image of group A, only AchE-positive area was stained; in the image of group B, myelin sheath was obscure; and in the image of group C, axons and myelin sheath could be indentified, the character of nerve fibers could be distinguished clearly and accurately, and the image segmentation of fascicles could be achieved easier than other 2 images. Conclusion The image of Karnovsky-Roots-toluidine blue-Ponceau 2R staining has no effect on the AchE-positive area in the image of Karnovsky-Roots staining and shows better texture feature. This improved histochemical process may provide ideal image for the three-dimensional reconstruction of peri pheral nerves.

Citation： LUO Peng,ZHANG Yi,QI Jian,ZHONG Yingchun,LIU Xiaolin.. RESEARCH OF HISTOCHEMICAL STAINING FOR IDENTIFYING THE FUNCTION AND MORPHOLOGY OF FASCICLES IN THREE-DIMENSIONAL RECONSTRUCTION OF PERIPHERAL NERVES. Chinese Journal of Reparative and Reconstructive Surgery, 2012, 26(4): 477-482. doi: Copy

1.	任高宏, 裴国献. 周围神经三维重建与可视化研究进展. 中国修复重建外科杂志, 2009, 23(2): 239-244.
2.	罗鹏, 刘小林, 戚剑, 等. 周围神经三维可视化模型建立中的功能束识别研究进展. 中华创伤骨科杂志, 2011, 13(2): 187-189.
3.	Sun K, Zhang J, Chen T, et al. Three-dimensional reconstruction and visualization of the median nerve from serial tissue sections. Microsurgery, 2009, 29(7): 573-577.
4.	张毅, 卢昊, 何彩风, 等. 周围神经功能束二维全景图像配准定位标志点的获取. 中国组织工程研究与临床康复, 2010, 14(35): 6567-6572.
5.	张毅, 刘小林, 钟映春, 等. 基于二次成像技术标志点获取和自动配准的长段周围神经功能束三维可视化关键技术研究. 中国修复重建外科杂志, 2010, 24(3): 325-331.
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7.	刘冬娟, 祝素文, 李艳杰. 硼砂甲苯胺蓝染液在半薄切片染色上的应用. 中国医科大学学报, 2004, 33(1): 33-33.
8.	田玉旺, 李丽. 神经髓鞘染色新方法的建立及其原理探讨. 中国组织化学与细胞化学杂志, 2004, 13(4): 543-545.
9.	许宗敬, 胡平. 显微图像纹理特征提取方法综述. 微计算机应用, 2009, 30(6): 6-13.
10.	印志鸿, 张季. 医学图像三维重建系统的关键技术研究与设计. 中国组织工程研究与临床康复, 2010, 14(43): 8036-8039.
11.	戚剑, 刘小林, 熊卓, 等. 短段腓总神经功能束三维重建的初步研究. 中国修复重建外科杂志, 2008, 22(9): 1031-1035.
12.	Kanaya F. Mixed nerve suture facilitated by enzyme-staining techniques. Tech Hand Up Extrem Surg, 2002, 6(3): 140-144.
13.	戚剑, 罗鹏, 程思红, 等. 基于图像分析的周围神经Karnovsky-Roots法染色规律的效果观察. 中国临床解剖学杂志, 2011, 29(4): 436-441.
14.	胥维勇, 杨群, 范小莉. 神经髓鞘染色及其应用. 中国组织化学与细胞化学杂志, 2004, 13(4): 546-547.
15.	方忠民. 医学图像的分割技术及其新进展. 长沙大学学报, 2008, 22(5): 94-97.
16.	栗卓, 吕广明, 刘苏, 等. 丽春红2R-亮绿法在中枢神经纤维束髓鞘染色中的应用. 交通医学, 2006, 20(5): 494-495.
17.	王俊杰, 刘家茂, 胡运发, 等. 图像拼接技术. 计算机科学, 2003, 30(6): 141-144.
18.	刘再涛, 魏本征, 柳澄. 一种基于视觉感知的复合医学图像分割算法. 郑州大学学报: 理学版, 2011, 43(1): 57-61.
19.	李芳, 钟映春, 张毅, 等. 神经切片图像中识别定位线断面的研究. 计算机工程与应用, 2010, 46(22): 203-205.
20.	钟映春, 戚剑, 刘小林, 等. 从图像中提取离散点状神经功能束边缘的研究. 系统仿真学报, 2011, 23(7): 1414-1418.

1. 任高宏, 裴国献. 周围神经三维重建与可视化研究进展. 中国修复重建外科杂志, 2009, 23(2): 239-244.
2. 罗鹏, 刘小林, 戚剑, 等. 周围神经三维可视化模型建立中的功能束识别研究进展. 中华创伤骨科杂志, 2011, 13(2): 187-189.
3. Sun K, Zhang J, Chen T, et al. Three-dimensional reconstruction and visualization of the median nerve from serial tissue sections. Microsurgery, 2009, 29(7): 573-577.
4. 张毅, 卢昊, 何彩风, 等. 周围神经功能束二维全景图像配准定位标志点的获取. 中国组织工程研究与临床康复, 2010, 14(35): 6567-6572.
5. 张毅, 刘小林, 钟映春, 等. 基于二次成像技术标志点获取和自动配准的长段周围神经功能束三维可视化关键技术研究. 中国修复重建外科杂志, 2010, 24(3): 325-331.
6. Zhang Y, Qi J, Liu XL, et al. Three-dimensional reconstruction of functional fascicular groups inside a segment of common peroneal nerve. Journal of Bioactive and Compatible Polymers, 2009, 24(1): 100-111.
7. 刘冬娟, 祝素文, 李艳杰. 硼砂甲苯胺蓝染液在半薄切片染色上的应用. 中国医科大学学报, 2004, 33(1): 33-33.
8. 田玉旺, 李丽. 神经髓鞘染色新方法的建立及其原理探讨. 中国组织化学与细胞化学杂志, 2004, 13(4): 543-545.
9. 许宗敬, 胡平. 显微图像纹理特征提取方法综述. 微计算机应用, 2009, 30(6): 6-13.
10. 印志鸿, 张季. 医学图像三维重建系统的关键技术研究与设计. 中国组织工程研究与临床康复, 2010, 14(43): 8036-8039.
11. 戚剑, 刘小林, 熊卓, 等. 短段腓总神经功能束三维重建的初步研究. 中国修复重建外科杂志, 2008, 22(9): 1031-1035.
12. Kanaya F. Mixed nerve suture facilitated by enzyme-staining techniques. Tech Hand Up Extrem Surg, 2002, 6(3): 140-144.
13. 戚剑, 罗鹏, 程思红, 等. 基于图像分析的周围神经Karnovsky-Roots法染色规律的效果观察. 中国临床解剖学杂志, 2011, 29(4): 436-441.
14. 胥维勇, 杨群, 范小莉. 神经髓鞘染色及其应用. 中国组织化学与细胞化学杂志, 2004, 13(4): 546-547.
15. 方忠民. 医学图像的分割技术及其新进展. 长沙大学学报, 2008, 22(5): 94-97.
16. 栗卓, 吕广明, 刘苏, 等. 丽春红2R-亮绿法在中枢神经纤维束髓鞘染色中的应用. 交通医学, 2006, 20(5): 494-495.
17. 王俊杰, 刘家茂, 胡运发, 等. 图像拼接技术. 计算机科学, 2003, 30(6): 141-144.
18. 刘再涛, 魏本征, 柳澄. 一种基于视觉感知的复合医学图像分割算法. 郑州大学学报: 理学版, 2011, 43(1): 57-61.
19. 李芳, 钟映春, 张毅, 等. 神经切片图像中识别定位线断面的研究. 计算机工程与应用, 2010, 46(22): 203-205.
20. 钟映春, 戚剑, 刘小林, 等. 从图像中提取离散点状神经功能束边缘的研究. 系统仿真学报, 2011, 23(7): 1414-1418.

Chinese Journal of Reparative and Reconstructive Surgery

RESEARCH OF HISTOCHEMICAL STAINING FOR IDENTIFYING THE FUNCTION AND MORPHOLOGY OF FASCICLES IN THREE-DIMENSIONAL RECONSTRUCTION OF PERIPHERAL NERVES

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