Microfluidic self-assembly of live Drosophila embryos for versatile high-throughput analysis of embryonic morphogenesis

被引:29
作者
Dagani, Gabriel T.
Monzo, Kate
Fakhoury, Jean R.
Chen, Chung-Chu
Sisson, John C.
Zhang, Xiaojing
机构
[1] Univ Texas, Dept Biomed Engn, Austin, TX 78712 USA
[2] Univ Texas, Microelect Res Ctr, Austin, TX 78712 USA
[3] Univ Texas, Sect Mol Cell & Dev Biol, Austin, TX 78712 USA
[4] Ind Technol Res Inst, Med Elect & Device Technol Ctr, Hsinchu, Taiwan
关键词
microfluidic; self-assembly; (polydimethylsiloxane) PDMS; Drosophila embryo; thermal perturbation; embryogenesis; cellularization;
D O I
10.1007/s10544-007-9077-z
中图分类号
R318 [生物医学工程];
学科分类号
0831 ;
摘要
A method for assembling Drosophila embryos in a microfluidic device was developed for studies of thermal perturbation of early embryonic development. Environmental perturbation is a complimentary method to injection of membrane-impermeable macromolecules for assaying genetic function and investigating robustness in complex biochemical networks. The development of a high throughput method for perturbing embryos would facilitate the isolation and mapping of signaling pathways. We immobilize Drosophila embryos inside a microfluidic device on minimal potential-energy wells created through surface modification, and thermally perturb these embryos using binary laminar flows of warm and cold solutions. We self-assemble embryos onto oil adhesive pads with an alcohol surfactant carrier fluid (detachment: 0.1 mL/min), and when the surfactant is removed, the embryo-oil adhesion increases to 2 similar to 5 mL/min flow rates, which allows for high velocities required for sharp gradients of thermal binary flows. The microfluidic thermal profile was numerically characterized by simulation and experimentally characterized by fluorescence thermometry. The effects of thermal perturbation were observed to induce abnormal morphogenetic movements in live embryos by using time-lapse differential interference contrast (DIC) microscopy.
引用
收藏
页码:681 / 694
页数:14
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