Measurement of cell migration in response to an evolving radial chemokine gradient triggered by a microvalve
Measurement of cell migration in response to an evolving radial chemokine gradient triggered by a microvalve
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DOI：
10.1039/b515560f
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发表时间：
2006-01-01
期刊：
影响因子：
6.1
通讯作者：
Folch, Albert
中科院分区：
文献类型：
--
作者：
Frevert, Charles W.;Boggy, Gregory;Folch, Albert
我们描述了一种基于微阀驱动的化学引诱剂从无细胞微室释放到含细胞微室的新型趋化性测定。将微瓣趋化装置 (mVCD) 放置在传统倒置显微镜的载物台上，以获得中性粒细胞在趋化因子 CXCL8/Interleukin-8 的径向对称演化梯度中迁移的延时显微照片。将荧光示踪剂添加到 CXCL8 溶液中以可视化梯度曲线的演变，以便在每个时间点的细胞位置都可以分配 CXCL8 浓度值。对单个中性粒细胞进行 90 分钟的跟踪表明，(a) 中性粒细胞迁移反应平均呈放射状指向 CXCL8 源； (b) 经常发生严重的非径向位移； (c)中性粒细胞群之间的迁移速度和方向存在相当大的异质性。使用定制的成像分析工具来提取迁移行为的测量值，例如速度、沿梯度径向轴的速度以及作为 CXCL8 浓度函数的转动角余弦。 mVCD 可以轻松地用于研究中性粒细胞以外的培养细胞的迁移行为。
We describe a novel chemotaxis assay based on the microvalve-actuated release of a chemoattractant from a cell-free microchamber into a cell-containing microchamber. The microvalve chemotaxis device (mVCD) was placed on the stage of a conventional inverted microscope to obtain time-lapse micrographs of neutrophils migrating in a radially-symmetric evolving gradient of the chemotactic factor CXCL8/Interleukin-8. A fluorescent tracer was added to the CXCL8 solution to visualize the evolution of the gradient profile, so that at each time point the cell positions could be assigned CXCL8 concentration values. Tracking of individual neutrophils for 90 minutes showed that (a) the neutrophil migratory response is, on average, radially directed towards the CXCL8 source; (b) significant non-radial displacements occur frequently; and (c) there is considerable heterogeneity in the migration speeds and directions amongst the neutrophil population. A custom-made imaging analysis tool was used to extract measurements of migratory behavior such as speed, velocity along the gradient's radial axis, and the cosine of the turning angle as a function of CXCL8 concentration. The mVCD can be easily adapted to study the migratory behavior of cultured cells other than neutrophils.