两种组织透明技术在免疫荧光染色观察脊髓3D结构中的应用

摘要

Objective To compare the application of CUBIC and iDISCO clearing methods in observing 3D imaging of spinal cord with immunofluorescent staining. Methods 1 mm thick spinal cord coronal sections were processed with CUBIC and iDISCO, respectively. The neurofilament (NF) protein was detected by immunofluorescent staining and then was observed by a laser confocal microscope. Results Compared with CUBIC, iDISCO had the advantages of shorter time, higher transparency (F=6.64, P<0.01), and deeper penetration (F=5117.55, P<0.01). Conclusion Immunofluorescent staining combined with iDISCO could completely observe the spinal axons with shorter time and better stain effect.%目的：观察CUBIC和iDISCO两种透明技术在脊髓免疫荧光染色中的效果。方法应用两种组织透明技术处理1 mm厚脊髓冠状切片，然后行神经丝(NF)蛋白免疫荧光染色，激光共聚焦扫描显微镜观察染色效果。结果与CUBIC透明相比，iDIS-CO透明技术所需时间短，透明度高(F=6.64, P<0.01)，免疫荧光染色深(F=5117.55, P<0.01)。结论应用iDISCO透明方法和免疫荧光染色相结合的方法可以完整观察脊髓轴突，更加简单快速，染色效果更好。