A microfluidic imaging chamber for the direct observation of chemotactic transmigration.
A microfluidic imaging chamber for the direct observation of chemotactic transmigration.
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DOI：
10.1007/s10544-010-9411-8
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发表时间：
2010-06
影响因子：
2.8
通讯作者：
Baskaran H
中科院分区：
文献类型：
--
作者：
Breckenridge MT;Egelhoff TT;Baskaran H
为了研究非肌肉肌球蛋白II(NM-II)在侵袭性细胞迁移中的作用，利用光刻和软光刻微制造技术设计和制造了微流控迁移小室。该室由两个通道组成，由垂直栅栏隔开，具有多个间隔的孔，宽度从6微米到16微米不等，长度从25微米到50微米不等。电池被电镀在栅栏一侧的通道中，而化学吸引剂则流经栅栏另一侧的通道。在这些小室中，可以用透射光或荧光光学观察到细胞，同时它们通过不同大小的毛孔进行化学作用，这些毛孔对转生施加了不同的机械阻力。作为对该装置的初步测试，我们比较了在NM-II抑制和不抑制NM-II的情况下，乳腺癌细胞通过不同孔径的趋化迁移。当细胞试图通过较小的毛孔拉出它们的细胞核时，观察到两种截然不同的速率，而较快的核传输模式严重依赖于NM-II运动活动。在单个实验系统中监测细胞通过不同尺寸的毛孔进行化学迁移的能力提供了关于毛孔大小如何影响细胞形态和迁移速度的新信息，提供了相对于其他体外迁移系统(如Boyden小室)的成像潜力的显著改善。
To study the roles of nonmuscle myosin II (NM-II) during invasive cell migration, microfluidic migration chambers have been designed and fabricated using photo- and soft-lithography microfabrication techniques. The chamber consists of two channels separated by a vertical barrier with multiple bays of pores with widths varying from 6 µm to 16 µm, and lengths varying from 25µm to 50µm. The cells are plated in the channel on one side of the barrier while a chemoattractant is flowed through the channel on the other side of the barrier. In these chambers, cells can be observed with transmitted light or fluorescence optics while they chemotax through various sized pores that impose differential mechanical resistance to transmigration. As an initial test of this device, we compared breast-cancer cell chemotactic transmigration through different pore sizes with and without inhibition of NM-II. Two distinct rates were observed as cells attempted to pull their nucleus through the smaller pores, and the faster nuclear transit mode was critically dependent on NM-II motor activity. The ability to monitor cells as they chemotax through pores of different dimensions within a single experimental system provides novel information on how pore size affects cell morphology and migration rate, providing a dramatic improvement of imaging potential relative to other in vitro transmigration systems such as Boyden chambers.
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影响因子：
7.5
作者：
Laemmermann, Tim;Sixt, Michael
通讯作者：
Sixt, Michael
影响因子：
6.1
作者：
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通讯作者：
Athanasiou, Kyriacos A.
影响因子：
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作者：
Laemmermann, Tim;Bader, Bernhard L.;Sixt, Michael
通讯作者：
Sixt, Michael
影响因子：
46.9
作者：
Jeon, NL;Baskaran, H;Toner, M
通讯作者：
Toner, M