网络讲座:量化成像分析流式细胞技术研究T细胞与APC细胞间相互作用


讲座主题:量化成像分析流式细胞技术研究T细胞与APC 细胞间相互作用
Title: Studies of T-cell/Antigen Presenting Cell Conjugation Using Imaging Flow Cytometry

内容摘要:
量化成像分析流式细胞技术是近年来广受国内外科学家关注的下一代流式细胞技术,它将流式细胞技术在免疫表型分析中的快速,灵敏以及统计学力量和量化成像在细胞功能及形态学研究中的细节展现能力完美结合,实现了细胞自噬、蛋白定位、细胞间相互作用、信号转导等多种热点研究的新的技术突破。本讲座将阐述量化成像分析流式细胞技术如何研究T细胞与APC细胞间相互作用。首先T细胞与APC之间要产生相互作用,需要形成细胞双粘体,利用量化参数可以检测T细胞与APC细胞双粘体比例;接下来信号分子会在细胞结合部位富集,以形成免疫突触。量化成像分析流式细胞技术可以评估细胞结合部位突触上信号分子LFA-1 and Lck的分布强度,以及与细胞活化密切相关的NFkB的核转位信息。
Abstract:
Imaging flow cytometry combines the speed, sensitivity, and phenotyping abilities of flow cytometry with the detailed imagery and functional insights of microscopy. This unique combination enables a broad range of applications that would be impossible using either technique alone. This seminar will explore two studies of the T-cell/Antigen Presenting Cell (APC) Immunological Synapse performed on FlowSight® and ImageStream®X Mark II imaging flow cytometers. In the first study we will demonstrate image-based parameters that were used to assess the frequency of T-cell/APC conjugates with an organized immunological synapse in an objective and statistically significant manner. In the second study we evaluate the specific location of the adhesion and signaling molecules LFA-1 and Lck within the immunological synapse complex in T-cells when presented with SEB, as well as the activation of the T cells in contact with APC by measuring the nuclear localization of NFkB in the T cell.

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