石蜡切片TUNEL法检测肉仔鸡法氏囊细胞凋亡的条件优化  

Optimization of Paraffin Section TUNEL Assay For Detecting Apoptosis in Broiler Bursa

作　　者：郭延锋[1] 李雪华[1] 付登峰[1] 胡建和[1,2] 王丽荣[1,2] 王三虎[1,2] 潘耀谦[1,2] 刘兴友[1,2] 

机构地区：[1]河南科技学院动物科学学院．河南新乡453003 [2]动物疫病和残留物防控河南省高校工程技术研究中心,河南新乡453003

出　　处：《动物医学进展》2010年第4期64-67,共4页Progress In Veterinary Medicine

基　　金：国家自然科学基金资助项目(30771601,30972202);河南省杰出人才创新基金项目(0521001800);河南省高校科技创新人才支持计划项目(2009HASTIT010)

摘　　要：从蛋白酶K的浓度、DAB显色时间、苏木素复染时间等对采用石蜡切片TUNEL法检测10日龄健康ROSS308肉仔鸡法氏囊组织内细胞凋亡的条件进行了优化。结果表明,TUNEL法最佳条件蛋白酶K浓度为20μg/mL,DAB显色时间为8 min,苏木素复染时间为10 s。优化后的石蜡切片TUNEL法,即60℃烤片5 h,20μg/mL蛋白酶K 37℃消化30 min,DAB显色8 min,苏木素复染10 s时,能特异地显示凋亡细胞为棕黄色,其他细胞核为蓝色,且对比明显,背景浅,组织结构完整清晰。To optimize the paraffin section TUNEL assay for detecting apoptosis in 10days healthy ROSS308 broiler bursa,the concentration of proteinase K,DAB chromogenic time and hematoxylin staining time were studied.The results showed the best conditions for TUNEL assay were 20 μg/mL concentration of proteinase K,8 min for DAB chromogenic time and 10 s for hematoxylin staining time.The optimized paraffin section TUNEL assay,which included 5 h for baking on 60 ℃,30 min for 20 μg/mL proteinase K digesting on 37 ℃,8 min for DAB chromogenic time and 10 s for hematoxylin staining time,specifically showed apoptotic cells in brown and other nuclei in blue.The comparison was obvious,the background was light,the structure was complete and clear,and was easy for micrograph.

关 键 词：TUNEL法 细胞凋亡 法氏囊 

分 类 号：S852.33[农业科学—基础兽医学]