苏丹黑B降低大鼠肠道自发荧光及其在免疫荧光染色中的应用

Sudan black B reducing the intestinal autofluorescence of rats and its application in immunofluorescence

目的探讨降低大鼠肠道组织自发荧光的简便可行方法。方法取正常SD大鼠(n=5)肠道做冷冻切片,荧光显微镜下观察自发荧光(AF),检测苏丹黑B(SBB)对AF的影响及其在肠道免疫荧光(IF)染色中的应用。结果大鼠各肠段组织在FITC和TRITC通道存在大量明显的AF,且不受血清封闭的影响;应用0.3%SBB处理组织10 min,可显著降低AF;IF染色加SBB处理后的肠道可见AF显著降低,目的蛋白的荧光表达不受影响。结论0.3%SBB染色10 min能显著降低大鼠肠道组织的AF,使IF特异性染色更加鲜明。

Objective To develop a feasible method to reduce the rat intestinal autofluorescence(AF).Methods Five rats were sacrificed.After cryostat section,intestinal AF was investigated by immunofluorescence(IF)before and after Sudan black B(SBB)treatment.Results The AF within the different rat intestinal segments was easily detected in both FITC-and TRITC-channel spectrum,which was not affected by serum blocking.After 0.3%SBB treatment for 10 minutes,a significant reduction of AF was observed in the rat intestinal tissue.Moreover,the SBB treatment did not affect the expression of CD45 in the rat intestine detected by IF staining,but reduced the AF remarkedly.Conclusion The AF in rat intestinal frozen section can be effectively reduced with 0.3%SBB treatment for 10 minutes.