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目的研究高效去除大鼠胸腺石蜡切片自发荧光的方法,并探索其在免疫荧光染色中的应用。方法应用硼氢化钠、台盼蓝、苏丹黑B对大鼠胸腺组织的石蜡切片进行处理并在荧光显微镜下观察自发荧光猝灭情况;并通过免疫荧光双标法对胸腺CD103及CD4阳性细胞进行染色探究其实用价值。结果经硼氢化钠处理后的切片红、绿、蓝三种颜色的自发荧光均较对照组明显减弱,且可增强胸腺组织切片免疫荧光染色效果。其他两种试剂处理后的石蜡切片自发荧光的强度与对照组无明显差异。结论硼氢化钠能显著的减低胸腺石蜡切片的自发荧光,可使免疫荧光特异性染色更加鲜明。
Objective To study the method for efficiently removing autofluorescence from rat thymus paraffin sections and to explore its application in immunofluorescence staining. Methods The paraffin sections of rat thymus were treated with sodium borohydride, trypan blue and Sudan black B, and the autofluorescence quenching was observed under a fluorescence microscope. The thymus CD103 and CD4 positive cells were immunofluorescently double labeled Dyeing to explore its practical value. Results After sodium borohydride treatment, the red, green and blue autofluorescence of the three colors were significantly weaker than the control group, and can enhance the immunofluorescence staining of thymus tissue sections. The intensity of autofluorescence of the paraffin sections treated with the other two reagents showed no significant difference from the control group. Conclusion Sodium borohydride can significantly reduce the autofluorescence of thymus paraffin sections and make immunofluorescence specific staining more distinct.