Karnovsky-Roots' stain of peripheral nerve based on image analysis
CU Jian, LUO Feng, CHENG Sai-Gong, LI Ceng-Hong, ZHANG Yi, LIU Xiao-Lin
Chinese Journal of Clinical Anatomy ›› 2011, Vol. 29 ›› Issue (4) : 436-441.
Karnovsky-Roots' stain of peripheral nerve based on image analysis
Objective To explore the feasibility of image analysis on the result of acetylcholinesterase stain (Karnovsky-Roots'method) of peripheral nerve, and provide the reference for the standard incubational conditions of the three-dimensional reconstruction of functional fascicles groups of peripheral nerves. Methods The samples came from fresh adult common peroneal nerve, which were serially horizontally sliced into 56 slices with the thickness of 10μm. All slices were randomly divided into seven groups with 8 slices in each group, including 1h, 2h, 4h, 8h, 12h, 24h and negative control group. After AChE histochemical staining, the stain characteristics of different nerve fiber in various stain phrases were observed under optic-microscope. The average area, density and integrated optical density (IOD) of AChE positive region of various stain period were analyzed by an image analytical software (Image-Pro Plus 6.0). T-test was adopted to distinguish the group differences through SPSS13.0. Results For the mean optical density value, there was notable diversity between 8h and 12h groups, but not for the 12h and 24h groups. The significant differences also appeared among 8h, 12h, and 24h groups for stain area and IOD. Furthermore, overstain was observed on slices of 24h group. Conclusions The staining results from 12h to 24h were stable and reliable, moreover, it is possible to identify different functional fascicles surrounding by perineurium. For three-dimensional reconstruction of functional fascicles groups of peripheral nerves, it can be taken as the standard stain incubational condition.
Acetylcholinesterase / Karnovsky-Roots / Peripheral nerve / Image analysis
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