Immunofluorescence method for stereoimaging of intrahepatic bile duct of child patient with biliary atresia

The invention discloses an immunofluorescence method for stereoimaging of intrahepatic bile ducts of children with biliary atresia. The immunofluorescence method comprises the following steps: fixing a specimen; an immunofluorescence method is improved, so that the intrahepatic bile duct can be marked; the liver tissue is transparentized by using a 3DISCO method, so that the liver tissue can maintain a fluorescence signal and achieve a transparentizing effect; and debugging the two-photon microscope, and selecting a proper optical filter for imaging. The method is mainly suitable for imaging of intrahepatic bile ducts (or intrahepatic blood vessels), and laser can smoothly penetrate through the tissues to carry out bile duct imaging by carrying out improved immunofluorescent staining and transparentizing treatment on the liver tissues. 本发明公开了一种胆道闭锁患儿肝内胆管立体成像的免疫荧光方法，包括：标本固定；改良免疫荧光方法使其能够对肝内胆管进行标记；利用3DISCO方法对肝脏组织进行透明化，使肝脏组织可保持荧光信号并达到透明化效果；调试双光子显微镜，选择合适的滤光片进行成像。本发明主要适用于肝内胆管(也可推及肝内血管)的成像，通过对肝脏组织进行改良免疫荧光染色及透明化处理，使激