探索双重荧光染色并利用两种不同显微镜在Hep-2细胞上观察抗核抗体荧光模式的临床价值  

Clinical Value of Observation of Fluorescence Pattern of Antinuclear Antibody in Hep-2 Cell by Using Double Staining Method and Two Different Microscopes

作　　者：王硕[1] 梁红艳[1] 姚玉虹[1] 迟伟群[1] 江跃红[1] 吕晓峰[1] 许铁[1] 于晓斐[1] 姜晓峰[1] 

机构地区：[1]哈尔滨医科大学附属第四医院检验科,黑龙江哈尔滨150001

出　　处：《标记免疫分析与临床》2015年第9期931-935,共5页Labeled Immunoassays and Clinical Medicine

基　　金：国家重大科学仪器设备开发专项(2011YQ040087)

摘　　要：目的探索荧光显微镜,激光扫描共聚焦显微镜以及双重荧光染色法在Hep-2细胞上观察抗核抗体荧光模式的临床价值。方法选取临床上抗核抗体呈阳性的血清样本结合到抗原片上,先后用异硫氰酸荧光素(FITC)和4',6-二脒基-2-苯基吲哚(DAPI)两种荧光染料孵育,之后在两种不同的显微镜下观察。结果 FITC荧光出现的部位是抗原抗体结合处,DAPI荧光出现的部位为细胞核,当通过DAPI荧光对细胞核定位后,可使得FITC的荧光模式更易于观察。本实验通过利用两种荧光标记的方法在两种显微镜上分别观察了斑点型、均质型、着丝点型、核点型、核膜型、胞浆颗粒型——抗高尔基体抗体。结论综合各种因素,利用双荧光染色法染色,并在激光扫描共聚焦显微镜下观察,对于在Hep-2细胞上观察抗核抗体荧光模式最具临床价值。Objective To evaluate the clinical value of fluorescence microscopy and confocal laser scanning microscope in detecting the fluorescence pattern of antinuclear antibody on Hep-2 cell Using double fluorescent labeling. Methods The samples whose antinuclear antibody was positive were chosen for the study. These samples were incubated with FITC and DAPI. Finally, fluorescence microscopy and confocal laser scanning microscope appeared on fluorescence were used to detect these samples respectively. the position of antigen-antibody binding site and Results The fluorescence of FITC and DAPI nucleus,respectively. It was easy to detect the pattern after locating the nucleus by the fluorescent of DAPI. The fluorescence patterns of Speckled Pattern, Homogeneous Pattern, Centromere Pattern, Nuclear Dot Pattern, Karyotheca Pattern, Cytoplasmic granules type were observed. Conclusion The double fluorescent labeling with the confocal laser scanning microscope to detect the fluorescence pattern of antinuclear antibody on Hep-2 cell is the most helpful method for clinical detection.

关 键 词：激光扫描共聚焦显微镜 荧光显微镜 抗核抗体 

分 类 号：R446.6[医药卫生—诊断学]