“小鼠脾脏单细胞悬液不同制备方法及其流式细胞术检测”综合实验的建立与探索  

Establishment and Preliminary Study of Comprehensive Experiment of“Different Preparation Methods and Flow Cytometry Detection of Mouse Spleen Single Cell Suspension”

作　　者：黄莹莹[1] 邢月婷 阿依西布·萨吾提 杨丹[1] 李艳伟[1] 王佳佳[1] 郭春 周南 HUANG Yingying;XING Yueting;Sawuti Ayixibu;YANG Dan;LI Yanwei;WANG Jiajia;GUO Chun;ZHOU Nan(Core Facilities,School of Medicine Zhejiang University,Hangzhou 310058,China)

机构地区：[1]浙江大学医学院公共技术平台,杭州310058

出　　处：《中国细胞生物学学报》2023年第4期680-687,共8页Chinese Journal of Cell Biology

基　　金：浙江大学实验技术研究项目(批准号:SYBJS202209)资助的课题。

摘　　要：由于流式细胞术应用中涉及的样品制备、仪器使用和数据解读均比较复杂,该文对流式实验教学的内容和形式进行了有益的探索,建立了“小鼠脾脏单细胞悬液不同制备方法及其流式细胞术检测”的综合性实验。新方法不仅包含小鼠原代组织取材、单细胞悬液不同制备方法(手动解离、自动解离)、流式细胞仪计数、显微观察等步骤,而且能涵盖流式抗体的配色与标记、流式细胞仪操作、数据分析(T淋巴细胞亚群的分群)等多个知识点,并围绕不同解离方法对实验结果的影响进行了探讨,极大地改善了实验教学效果和质量,充分发挥了流式细胞术在高校教学和科研中的重要支撑作用,使学生学会完整的流式技术链条,并提升了其科研能力与创造力,从而为培养复合型科研人才提供助力。Due to the complexity of sample preparation,instrument manipulation and data analysis involved in FCM(flow cytometry)applications,beneficial explorations on the content and form of FCM experimental teaching were conducted.Accordingly,a comprehensive experiment of“different preparation methods and FCM detection of mouse spleen single cell suspension”has been established.The new method can not only include sampling of mouse primary tissue,different preparation methods of single-cell suspension(manual dissociation,automatic dissociation),flow cytometer counting,microscopic observation,etc.,but also cover many knowledge points,such as color matching and labeling of antibodies in FCM,operation of flow cytometer,data analysis(cluster of T lymphocyte subsets),etc.The influences of different dissociation methods on the experimental results were discussed.It greatly improves the effectiveness and quality of experimental teaching,gives full play to the important supporting role of FCM in teaching and scientific research in colleges and universities.Based on these lessons,students can learn the complete technology chain of FCM,which enhances their scientific research ability and creativity.The comprehensive experiment would provide impetus for the cultivation of compound scientific research talents.

关 键 词：单细胞悬液 手动解离 自动解离 流式检测 实验教学 

分 类 号：Q95-33[生物学—动物学]