Kress Holger, Park Jin-Gyu, Mejean Cecile O, Forster Jason D, Park Jason, Walse Spencer S, Zhang Yong, Wu Dianqing, Weiner Orion D, Fahmy Tarek M, Dufresne Eric R
Department of Mechanical Engineering, Yale University, New Haven, Connecticut, USA.
Nat Methods. 2009 Dec;6(12):905-9. doi: 10.1038/nmeth.1400. Epub 2009 Nov 15.
Molecular gradients are important for various biological processes including the polarization of tissues and cells during embryogenesis and chemotaxis. Investigations of these phenomena require control over the chemical microenvironment of cells. We present a technique to set up molecular concentration patterns that are chemically, spatially and temporally flexible. Our strategy uses optically manipulated microsources, which steadily release molecules. Our technique enables the control of molecular concentrations over length scales down to about 1 microm and timescales from fractions of a second to an hour. We demonstrate this technique by manipulating the motility of single human neutrophils. We induced directed cell polarization and migration with microsources loaded with the chemoattractant formyl-methionine-leucine-phenylalanine. Furthermore, we triggered highly localized retraction of lamellipodia and redirection of polarization and migration with microsources releasing cytochalasin D, an inhibitor of actin polymerization.
分子梯度对于各种生物过程都很重要,包括胚胎发育和趋化作用期间组织和细胞的极化。对这些现象的研究需要控制细胞的化学微环境。我们提出了一种建立分子浓度模式的技术,该模式在化学、空间和时间上具有灵活性。我们的策略使用光学操纵的微源,其能稳定地释放分子。我们的技术能够在低至约1微米的长度尺度以及从几分之一秒到一小时的时间尺度上控制分子浓度。我们通过操纵单个人类中性粒细胞的运动性来演示这项技术。我们用装载有趋化因子甲酰甲硫氨酸-亮氨酸-苯丙氨酸的微源诱导了细胞的定向极化和迁移。此外,我们用释放细胞松弛素D(一种肌动蛋白聚合抑制剂)的微源触发了片状伪足的高度局部收缩以及极化和迁移方向的重新定向。