An automated multi well cell track system to study leukocyte migration
An automated multi well cell track system to study leukocyte migration
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DOI：
10.1016/s0022-1759(03)00262-x
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发表时间：
2003-09-01
影响因子：
2.2
通讯作者：
Boezeman, JBM
中科院分区：
文献类型：
--
作者：
Krooshoop, DJEB;Torensma, R;Boezeman, JBM
设计自动图像处理系统来确定单个细胞的迁移特征不是微不足道的。每个测试样品都需要单独记录，并且通过延时显微镜在二维或三维迁移系统中分析单个细胞轨迹是非常费力的。在这里，我们描述了一个新的自动细胞跟踪系统（ACTS）。除了现有分析系统使用的对比度差异之外，ACTS算法还基于连续图像中的形态学相似性来识别细胞，并适应单个细胞在迁移过程中的连续形状变化。该系统便于在一次实验中同时分析多个细胞和测量多个孔。我们在涂有ICAM-1-Fc蛋白或对照CD14-Fc蛋白的标准96孔微滴板上验证了该系统，研究了HSB-2 T细胞的迁移。HSB-2 T细胞在ICAM-1-Fc上的迁移是由白细胞功能相关抗原-1 （LFA-1）介导的，迁移细胞的数量和速度取决于ICAM-1-Fc的浓度。我们展示了迁移数据的自动化分析产生了与手工分析相似的结果，但是所用的时间很短。我们得出结论，该系统非常适合于精确监测单个细胞的迁移行为。多孔并行分析使这种设置适合于高通量筛选，其中同时测试多种组分对细胞迁移的影响。(C) 2003 Elsevier B.V.版权所有
Design of automated image processing systems to determine migration characteristics of individual cells is not trivial. Every test sample requires separate recording and the analysis of individual cell tracks in two- or three-dimensional migration systems by time-lapse microscopy is extremely laborious. Here, we describe a new Automated Cell Track System (ACTS). In addition to contrast differences, which are used by existing analysis systems, the ACTS algorithms recognize cells on the basis of morphological similarities in successive images and adapt to the continuous shape changes of individual cells during migration. The system facilitates simultaneous analysis of multiple cells and the measurement of multiple wells in one single experiment.We validated the system studying HSB-2 T cell migration in standard 96-well microtiter plates coated with ICAM-1-Fc protein or control CD14-Fc protein. Migration of HSB-2 T cells on ICAM-1-Fc is Leukocyte Function-associated Antigen-1 (LFA-1)-mediated and both the number and the speed of migrating cells depend on the ICAM-1-Fc concentration.We show that automated analysis of the migration data yields similar results as manual analysis, but in a fraction of the time. We conclude that this system is extremely well suited to precisely monitor the migratory behavior of individual cells. The analysis of multiple wells in parallel makes this set-up appropriate in high throughput screening in which multiple components are simultaneously tested for their effect on cell migration. (C) 2003 Elsevier B.V. All rights reserved.