Dagani Gabriel T, Monzo Kate, Fakhoury Jean R, Chen Chung-Chu, Sisson John C, Zhang Xiaojing
Department of Biomedical Engineering, The University of Texas at Austin, 1 University Station, ENS 12, Austin, TX 78712-0238, USA.
Biomed Microdevices. 2007 Oct;9(5):681-94. doi: 10.1007/s10544-007-9077-z.
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 approximately 25 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.
为了研究早期胚胎发育的热扰动，开发了一种在微流控装置中组装果蝇胚胎的方法。环境扰动是一种补充方法，可用于注射不透膜的大分子以测定基因功能和研究复杂生化网络中的稳健性。开发一种高通量的胚胎扰动方法将有助于信号通路的分离和映射。我们将果蝇胚胎固定在微流控装置内通过表面改性创建的最小势能阱中，并使用温热和冷溶液的二元层流对这些胚胎进行热扰动。我们使用酒精表面活性剂载液（分离速度：0.1 mL/分钟）将胚胎自组装到油粘性垫上，当去除表面活性剂时，胚胎与油的粘附力增加到约25 mL/分钟的流速，这允许热二元流的陡峭梯度所需的高速度。微流控热分布通过模拟进行数值表征，并通过荧光测温法进行实验表征。通过使用延时差分干涉对比（DIC）显微镜观察到热扰动的影响会在活胚胎中诱导异常的形态发生运动。
