同一细胞非放射性双原位杂交信号显示技术

Demonstration of non-radioactive in situ hybridization with double signals in the single cell

目的 :探讨同一细胞内双原位杂交信号的显示技术。方法 :以地高辛标记的polβ探针加生物素标记的表皮生长因子受体 (EGFR)探针或生物素标记的wtp5 3探针对人食管癌Eca 10 9细胞同时进行原位杂交后 ,以anti Dig AP孵育 ,以AP Red为底物显色 ,杂交信号呈桔红色或桔黄色细颗粒。继之以SA AP孵育后以NBT/BCIP底物显色 ,杂交信号呈蓝紫色颗粒。结果 :在同一细胞内可见桔黄色和蓝紫色颗粒双杂交信号。结论 :应用非放射性双杂交信号的原位杂交技术可观察单个细胞内

Aim:To detect how the double hybridization signals co expressed in the single cell. Methods: The in situ hybridization with Dig labeled polβ probe combined with Bio labeled EGFR probe or wtp53 probe in Eca 109 cells, and incubation with anti Dig AP were performed.The first signals appeared orange yellow granules after the AP Red was used as the substrate. Subsequently, the second signals showed bluish violet granules after incubation with SA AP and NBT/BCIP used as the substrate.Results: The double signals in orange yellow mixed with bluish violet colors were found in the same cells.Conclusion: The gene co expression could be detected and localized in the single cell by in situ hybridization technique with double signals.