CdTe纳米晶标记的Annexin V抗体识别大鼠感觉神经的实验研究

The experimental study of sensory nerve fascicles identification with Annexin V antibody labeled by CdTe nanocrystals

目的研究CdTe纳米晶标记的Annexin V抗体在鉴别大鼠周围神经功能束的效果。方法将10只Vista大鼠随机分成两组,每组5只,一组为实验组,另一组为实验对照组。将大鼠处死后,取一侧坐骨神经干及其肌支和腓肠神经,立即做快速冰冻切片,实验组以CdTe纳米晶标记的Annexin V抗体做为孵育液,对照组以正常羊血清代替抗体,在37℃恒温箱中孵育,分别在10 min,15 min,20 min及30 min后取出,以0.01 mol/L的PBS洗涤5min后在荧光显微镜下观察。结果对照组在镜下未发现荧光,实验组坐骨神经肌支在各时间段均未发现阳性染色,坐骨神经干感觉束支及腓肠神经在10 min时出现阳性染色,但难以清晰地显示出完整的神经束,在15 min时可清晰地见到神经束,而20min和30 min时荧光强度与15 min时差别不大。结论应用CdTe纳米晶标记的Annexin V抗体可快速识别大鼠感觉神经,可为临床鉴别周围神经束性质提供一种新的方法。

Objective To study the effect of Annexin V antibody labeledby CdTe nanocrystals in differentiation peripheral nerve fascicles. Methods Ten rats were divided into two groups randomly, 5 each group. Excise sural nerve, sciatic nerve and its muscle branches after the rats killed,tile excised nerve stumps were quick frozen and cross-sectioned （20 tan） with a cryostat microtome.The experimental group use Annexin V antibody/abeled by CdTe nanocrystals as incubating liquid, the controlled use CdTe nanocrystals, then the specimens were incubated in 37℃ incubator. Taken out to observe under fluorescent microscop at 10 min, 15 min,20 min and 30 min respectively. Results The fluorescence was not seen in controlled group, while in experimental group, different from the sensory branches of sciatic nerve and sural nerve, the muscle branches of sciatic nerve could not see thefluorescence, the fluorescence intensity at 10 min was not enough to show the nerve fascicles clearly,we could see the fascicles with red fluorescence clearly at 15 min, the fluorescence intensity at 20 min and 30 min had httie difference with that at 15 min. Conclusion The Annexin V antibody labeled by CdTe nanocrystals can identify sensory nerve fascicles of rat rappidly, it is possible to provide a new method for clinical differentiation peripheral nerve nerve fascicles.