摘要
目的:使用高灵敏图像处理软件Volocity分析T细胞和树突状细胞动态相互作用。方法:将磁珠分选后的CD3+T细胞和流式细胞仪分选后CD11c+MHCⅡ+的树突状细胞进行荧光标记,采用激光共聚焦显微镜在二维和三维水平成像后,使用Volocity软件分析两种细胞的动态相互作用。结果:使用Volocity软件分析了视野内T细胞和树突状细胞形态、接触体积和运动轨迹的变化趋势,统计了细胞接触时间,并比较手动分析和软件分析细胞运动轨迹的差异,显示两种细胞在30 min内发生了明显接触,促进后续免疫反应发生。结论:使用Volocity系统分析T细胞和树突状细胞动态相互作用,建立了两种细胞动态相互作用的方法。
Objective: To analyze dynamic interaction of T cells and dendritic cells by using Volocity high performance image analysis software. Methods: The CD3+T cells were isolated via MACS, CD11c+MHCⅡ+ dendritic cells were purified by flow cytometry, Then the two cells were labeled and analyzed using the Perkin Elmer laser confocal microscopy imaging in 2D and 3D level. The dynamic interaction between the two kinds of cells was analyzed by Volocity image analysis software. Results: Analysis of the dynamic interaction parameters include: morphological changes, cell tracking, the contact time, the contact area, classification of contacts and so on, the difference between the manual and software analysis of cell tracking were counted. In this experiment, the interactions between dendritic cell and T cells have been visualized in real time. The contact leads to formation of immunological synapses at the interface between the cells. Thesynapse is divided into close contact and loose contact. There were significant changes in cell morphology, cell became more irregular. In addition, the tracking of manual and volocity software analysis of cells has very significant difference. The contact between the two types of cells promote that an immune response is launched. Conclusion: This method to study dynamic interaction between T cells and dendritic cell using Volocity software were built successfully.
出处
《中国医学装备》
2015年第2期1-5,共5页
China Medical Equipment
基金
国家自然科学基金(81273217)"Foxp1对调节性T细胞和自身免疫病的调控作用"