显微成像分析技术在中性粒细胞运动及吞噬功能研究中的应用

发布时间：2018-03-13 23:35

本文选题：显微成像技术　切入点：图像定量分析　出处：《中国实验血液学杂志》2015年03期 　论文类型：期刊论文

【摘要】：目的:使用转盘式激光共聚焦显微镜及显微图像处理软件对中性粒细胞的运动及吞噬功能进行分析与评估。方法:使用Percoll分离液分离小鼠骨髓中性粒细胞,用PE荧光标记后与FITC标记的酵母聚糖颗粒(Zymosan)混匀,置于转盘式激光共聚焦显微镜下进行多通道时间序列成像。使用Volocity和Image J分析中性粒细胞的运动及吞噬相关参数,包括形态变化、运动轨迹、伪足振荡、结合吞噬指数,得出时间序列下各项参数的变化趋势。结果:中性粒细胞在受到Zymosan诱导后趋化作用明显,大部分细胞在短时间内发生极性化,在40分钟内能快速结合Zymosan,行使吞噬功能。结论:使用显微成像技术结合图像处理分析技术可准确、定量地分析中性粒细胞运动及吞噬功能,直观快速地对中性粒细胞的生理学状态进行初步评估。通过这项技术,得到正常生理状态下中性粒细胞对Zymosan的快速响应情况,为今后中性粒细胞相关功能学研究提供了参考依据。
[Abstract]:Objective: to analyze and evaluate the movement and phagocytosis of neutrophils by using rotating disc laser confocal microscope and microimage processing software. After being labeled with PE fluorescence and mixed with FITC labeled yeast polysaccharide granules, the multichannel time series imaging was performed under a rotating disc laser confocal microscope. The parameters of neutrophil motion and phagocytosis were analyzed by Volocity and Image J. Including morphological changes, motion trajectories, pseudopod oscillations, combined with phagocytosis index, the change trend of parameters under time series was obtained. Results: neutrophil chemotaxis was obvious after induced by Zymosan. Most of the cells were polarity in a short time and could combine with Zymosanin in 40 minutes to perform phagocytosis. Conclusion: microimaging combined with image processing can accurately and quantitatively analyze the function of neutrophil motion and phagocytosis. The physiological state of neutrophils was evaluated intuitively and quickly. The rapid response of neutrophils to Zymosan in normal physiological state was obtained by this technique. It provides a reference for the study of neutrophil related function in the future.
【作者单位】：中国医学科学院北京协和医学院血液病医院血液学研究所实验血液学国家重点实验室;
【基金】：国家自然基金(81300436,81470280) 实验血液学国家重点实验室自由申请项目(ZZ13-05)
【分类号】：R392

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