原位杂交和免疫组化双标记技术在大鼠全脑脑片培养中的应用

Application of Double Staining of In Situ Hybridization and Immunohistochemistry in Brain Slices

目的探讨联合应用原位杂交(in situ hybridization,ISH)和免疫组化(immunohistochemistry,IHC)技术检测全脑脑片特定核酸和蛋白的表达。方法在静态全脑脑片培养的基础上,将培养的脑片进行冰冻切片,然后联合应用原位杂交和免疫组化技术检测特定基因的表达情况。结果脑片冰冻切片的质量可以进行后续的实验;与单纯应用一种方法相比,联合应用两种技术可同时在核酸水平和蛋白水平检测待测基因,两种显色方法不相互冲突,且可以更直观地显示核酸和蛋白的表达模式。结论原位杂交和免疫组化双染技术明显优于单纯的原位杂交技术或免疫组化;该双染技术可用于脑片培养中的基因和蛋白检测。

Objective To investigate the use of immunohistochemistry（IHC） in combination with in situ hybridization（ISH） to detect a special gene at nucleotide level and protein level at the same time on whole brain slices. Methods After culturing the whole brain slices of rats,HE staining was applied to assess the quality of frozen section of brain slices. Then ISH was used in combination with IHC to detect the expression of a special gene. Results Whole brain slices grew well enough to be used in following research. Compared to either IHC or ISH,combination of both was able to detect the gene at nucleotide level and protein level at the same time. Conclusion Double staining was superior to IHC or ISH. This technique could be used in detecting the nucleotide and protein in brain slices.