受体光漂白FRET技术的实践与优化  

Practice and optimization of acceptor photobleaching FRET technology

作　　者：刘双双[1] 王君 尹伟[1] 娄绘芳[1] LIU Shuangshuang;WANG Jun;YIN Wei;LOU Huifang(Core Facilities of School of Medicine,Zhejiang University,Hangzhou 310058,China;Zhejiang University Hospital,Zhejiang University,Hangzhou 310058,China)

机构地区：[1]浙江大学医学院公共技术平台,浙江杭州310058 [2]浙江大学浙江大学校医院,浙江杭州310058

出　　处：《实验技术与管理》2020年第12期39-43,共5页Experimental Technology and Management

基　　金：浙江省基础公益研究计划项目(LGF20H090012)。

摘　　要：蛋白质之间的相互作用对生命活动的调节具有重要意义,为了精确检测纳米级蛋白质之间的相互作用,促进高端成像技术的发展。利用共聚焦显微镜系统和图像处理软件ImageJ,开发和优化了受体光漂白荧光共振能量转移(FRET)技术。该实验总结了FRET荧光对的特点并提出选择方法,确定了受体光漂白FRET技术的操作步骤,分析了FRET技术的限制条件并给出了优化方案,详细描述了图像展示和数据分析的具体方法。该文建立了从实验设计到数据分析的一整套实用可靠的操作方案,可为显微成像技术从业者提供借鉴。The interaction between proteins is of great significance to the regulation of life activities.In order to accurately detect the interactions between nanoscale proteins and promote the development of high-end imaging technology,the confocal microscope system and image processing software ImageJ are used to develop and optimize the acceptor photobleaching fluorescence resonance energy transfer(FRET)technology.This experiment has summarized the characteristics of FRET fluorescence pairs,proposed a selection method,determined the operating steps of acceptor photobleaching FRET technology,analyzed the limitations of the FRET technology and gave an optimization plan,and described the specific methods of image display and data analysis in detail.A set of practical and reliable operation schemes from experimental design to data analysis is established to provide reference for practitioners of microscopic imaging technology.

关 键 词：荧光共振能量转移 供体-受体对 蛋白质相互作用 

分 类 号：Q-336[生物学]