激光共聚焦扫描显微镜观测自噬小体变化的应用  

Application of Laser Confocal Scanning Microscope in Observing Autophagy

作　　者：张冰娜[1] 叶丹彦 张丹桂 李璐 杨旅军[1] ZHANG Bingna;YE Danyan;ZHANG Dangui;LI Lu;YANG Lvjun(Research Center of Translational Medicine,Second Affiliated Hospital of Shantou University Medical College,Shantou 515041,Guangdong,China)

机构地区：[1]汕头大学医学院第二附属医院转化医学研究中心,广东汕头515041

出　　处：《汕头大学学报（自然科学版）》2021年第1期76-80,F0003,共6页Journal of Shantou University：Natural Science Edition

基　　金：高水平大学建设计划临床医学重点建设学科专项资金(粤教科函(2018)181号)。

摘　　要：文章探讨激光共聚焦扫描显微镜技术在检测低氧诱导小鼠心肌细胞损伤自噬小体变化的应用价值.在无菌条件下,取胚胎型小鼠心肌细胞备用.采用低氧环境+缺氧液方法诱导心肌细胞损伤并发生自噬现象作为实验组,正常环境下培养细胞的方法作为对照组.待心肌细胞长到一定程度后,分别利用激光共聚焦扫描显微镜和正置荧光显微镜对免疫荧光染色的自噬小体表达的LC3A/B蛋白情况进行观测并比较2种检测方法的优缺点.结果显示利用低氧环境+缺氧液方法诱导小鼠心肌细胞损伤并自噬小体形成是可行的.与正常对照组比较,低氧缺氧诱导小鼠心肌细胞损伤组(实验组)自噬小体呈高表达状态.与普通型正置荧光显微镜比较,激光共聚焦扫描显微镜成像速度快,一次可以获得多副图像且图像清晰.激光共聚焦扫描显微镜技术具有成像速度快、灵敏度高和精确度高等优点,可以实时、准确观测低氧诱导小鼠心肌细胞损失自噬小体的表达情况,具有重要临床应用价值.To investigate the application value of laser confocal scanning microscopy(LSCM)in detecting the changes of autophagy bodies in hypoxic-induced myocardial injury in mice.Under aseptic condition,cardiomyocytes of embryonic type mice were taken for standby.The experimental group was treated with hypoxic environment and hypoxia solution to induce myocardial cell injury and autophagy,and the control group was cultured in normal environment.After the cardiomyocytes grew to a certain extent,the LC3A/B protein expression of autophagosome was observed by laser confocal scanning microscope and forward fluorescence microscope,respectively.The advantages and disadvantages of the two methods were also compared.It is feasible to induce myocardial cell injury and autophagy in mice by hypoxia environment and hypoxia solution.Compared with the normal control group,the autophagy of the mice myocardial cell injury group(experimental group)was highly expressed.Compared with the normal fluorescent microscope,the laser confocal scanning microscope has a fast imaging speed,and can obtain multiple images at a time and the image is clear.Laser confocal scanning microscope has the advantages of fast imaging speed,high sensitivity and high accuracy.It can be used to observe the expression of autophagy in mice cardiomyocytes induced by hypoxia in real time and accurately,and has important clinical application value.

关 键 词：激光共聚焦扫描显微镜 正置荧光显微镜 自噬小体 

分 类 号：TN24[电子电信—物理电子学]