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目的观察荧光标记物DiI在已固定的人胚胎周围神经中的示踪显影,探讨DiI在周围神经示踪中的应用可行性。方法利用人胚胎新鲜标本(非疾病死亡的引产胎儿),胎龄>20w,经升主动脉灌注10%福尔马林进行固定,并浸泡于固定液8~10d后,将微量的DiI结晶分别植入人胚胎标本三叉神经的眼神经、上颌神经、下颌神经及三叉神经根中,放入固定液在37℃恒温箱内贮存3个月,经冰冻切片,置荧光显微镜下观察评价示踪效果。结果 DiI注入眼神经、上颌神经及下颌神经中后,可在三叉神经节内观察到许多逆行标记的神经节细胞和纤维,且细胞及细胞突起显示较好;DiI注入三叉神经根中后,三叉神经节内也可见许多顺行标记的神经节细胞和纤维,细胞及细胞突起也显示较好。结论 DiI是一种良好的荧光类双向神经示踪剂,用于已固定人胚胎周围神经束路的失踪研究是可行的。
Objective To observe the tracer imaging of fluorescent labeled DiI in fixed human peripheral nerves and to explore the feasibility of using DiI in peripheral nerve tracing. Methods Fresh fetal human embryos (induced fetus without disease death), gestational age> 20w, fixed by perfusion of 10% formalin in ascending aorta and immersed in fixative solution for 8 ~ 10d, Implantation of human embryonic specimens of the trigeminal nerve in the oculomotor nerve, maxillary nerve, mandibular nerve and trigeminal nerve root, into the fixed fluid stored at 37 ℃ incubator for 3 months, frozen section, under the fluorescence microscope to evaluate the tracer evaluation . Results After DiI injection into the oculomotor nerve, maxillary nerve and mandibular nerve, many retrograde labeled ganglion cells and fibers were observed in the trigeminal ganglion, and the cells and cells showed better projections. After DiI was injected into the trigeminal nerve root, Numerous ganglion-labeled ganglion cells and fibers were also found in the ganglia, and cell and cell processes also showed better. Conclusion DiI is a good fluorescence bi-directional neurotracer. It is feasible to study the disappearance of the peripheral nerve bundles in human embryos.