Nanolitre liquid patterning in aqueous environments for spatially defined reagent delivery to mammalian cells.
Nanolitre liquid patterning in aqueous environments for spatially defined reagent delivery to mammalian cells.
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作者：
微尺度生物调控技术可以调节细胞-材料的相互作用和细胞形状，并能够以细胞和试剂高效的方式进行多路高通量研究。大多数可用的技术依赖于印章、大头针或口罩与主要是干燥表面的物理接触。喷墨和压电打印以非接触方式执行，但仍需要基本干燥的衬底以确保打印图案的保真度。因此，这些现有的方法仅限于在活细胞的微妙表面上形成图案，因为物理接触或基本干燥的条件会对它们造成损害。具有层流的微流控图案化确实允许在全水环境中进行非接触式图案化，但吞吐量和试剂在界面流中的扩散有限。在这里，我们描述了一种聚合物双水相系统(ATPS)，该系统能够在覆盖细胞单分子层的第二水相中图案化任意形状的含试剂水相的纳米升。在适当的介质配方下，感兴趣的试剂仍然局限在图案化的相中，没有明显的扩散。全水环境确保了较高的试剂活性和细胞活力。这一策略的实用性通过将遗传物质模式化地输送到哺乳动物细胞来进行基因表达和基因沉默的表型筛选而得到证明。
Microscale biopatterning allows regulation of cell-material interactions and cell shape, and enables multiplexed high throughput studies in a cell and reagent efficient manner. The majority of available techniques rely on physical contact of a stamp, pin, or mask with mainly a dry surface. Inkjet and piezoelectric printing is performed in a non-contact manner but still requires a substantially dry substrate to ensure fidelity of printed patterns. These existing methods, therefore, are limited for patterning onto delicate surfaces of living cells because physical contact or substantially dry conditions are damaging to them. Microfluidic patterning with laminar streams does allow non-contact patterning in fully aqueous environments but with limited throughput and reagent diffusion across interfacial flows. Here, we describe a polymeric aqueous two-phase system (ATPS) that enables patterning nanoliters of a reagent-containing aqueous phase, in arbitrary shapes, within a second aqueous phase covering a cell monolayer. With the appropriate media formulation, reagents of interest remain confined to the patterned phase without significant diffusion. The fully aqueous environment ensures high reagent activity and cell viability. Utility of this strategy is demonstrated with patterned delivery of genetic materials to mammalian cells for phenotypic screening of gene expression and gene silencing.
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影响因子：
3.3
作者：
Li, Xiao-Yan;Ota, Ichiro;Weiss, Stephen J.
通讯作者：
Weiss, Stephen J.
影响因子：
5.2
作者：
Chen, Silvia S.;Fitzgerald, Wendy;Margolis, Leonid
通讯作者：
Margolis, Leonid
影响因子：
7.8
作者：
STREULI, CH;SCHMIDHAUSER, C;DERYNCK, R
通讯作者：
DERYNCK, R
DOI：
10.1083/jcb.149.6.1309
发表时间：
2000-06-12
期刊：
The Journal of cell biology
影响因子：
--
作者：
Hotary K;Allen E;Punturieri A;Yana I;Weiss SJ
通讯作者：
Weiss SJ
影响因子：
14
作者：
Roth, EA;Xu, T;Boland, T
通讯作者：
Boland, T