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Microchannel arrays with improved accessibility and use for cell studies and emulsification

机译：具有改进的可及性的微通道阵列，可用于细胞研究和乳化

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Abstract: Arrays of microgrooves (groove width; 2, 3, 4, 5, 6, 7, 8, 10, 12, and 14 micrometer, groove interval; width x3, x10, and x20, one size and interval per chip) each connecting a center well and a side edge of a silicon substrate were created by photolithography and anisotropic wet etching. A penetrating hole was made by sand blast at the substrate center for the access to the center well. By tightly covering the substrate surface with a glass plate, the microgroove arrays were converted to microchannel arrays having one ends open at the side edges of the substrate. These microchannel arrays were used for cell trapping for microinjection and also used for emulsification. Poplar (Populus alba) protoplasts were used for the test of cell trapping. Cells showed a very large variation in size and irregularity in shape, and, furthermore, the protoplast preparation contained a number of cell membrane fragments and chloroplasts. Despite the cell size and shape variations and obstruction by the admixtures, many cells could be trapped by aspiration at the channel ends because of their openness to the outside free space and also their large multiplicity in parallel. The free space outside the side of the substrate allowed a free manipulation of a glass micropipette under microscopic observation using transmitted illumination. The microscopic observation direction nearly perpendicular to the movement directions of the micropipette further allowed the movement of the pipette tip nearly always in focus. These led to an easy pointing and puncturing. In addition, the cell trapping points in a line made successive approach to adjacent cells easier. Soybean oil containing 1.5 wt% polyoxyethylene(20)sorbitan monoolete as a surfactant was forced to flow into physiological saline filling the outside of the substrate through the microchannels. Regularly sized oil particles were created by this process with a variation coefficient (S.D./mean) 16% of their diameter. This variation, which is larger than those (minimum 1 - 2%) obtained in our previous trials using our previous microchannel arrays, appeared to be attributable to an irregularity of the channel ends due to microchipping by saw cutting. As an advantage over the previous ones, the present microchannel arrays allowed an easy collection of the created oil particles and also an easy change of the composition of the suspending fluid during the process. The substrate side surface is thus indicated to be useful for interfacing structures or devices microfabricated in the main substrate surface, which may be covered with a glass plate, with conventional or hand-operated tools or processes outside the substrate. !16
机译：摘要：每个微槽阵列（槽宽； 2、3、4、5、6、7、8、10、12和14微米，凹槽间隔；宽度x3，x10和x20，每个芯片一个尺寸和间隔）通过光刻和各向异性湿法蚀刻形成连接中心阱和硅衬底的侧边缘的连接。通过喷砂在基片中心开一个通孔，以通向中心孔。通过用玻璃板紧密覆盖基板表面，将微槽阵列转换成在基板的侧边缘具有一端开口的微通道阵列。这些微通道阵列用于细胞捕获以进行显微注射，也用于乳化。杨树（Populus alba）原生质体用于细胞捕获测试。细胞显示出很大的尺寸变化和形状不规则，此外，原生质体制剂包含许多细胞膜碎片和叶绿体。尽管细胞大小和形状变化以及混合物的阻碍，许多细胞仍可能由于对外部自由空间的开放性以及它们平行的大量多样性而被通道末端的抽吸吸引。基板侧面外侧的自由空间允许在使用透射光的显微镜观察下自由操作玻璃微量移液器。几乎垂直于微量移液器的运动方向的微观观察方向进一步允许移液器尖端的运动几乎总是聚焦。这些导致了轻松的指向和穿孔。另外，一条线中的细胞捕获点使相邻细胞的连续接近变得更容易。含有1.5 wt％的聚氧乙烯（20）脱水山梨糖醇单油酸酯作为表面活性剂的大豆油被迫通过微通道流入充满生理盐水的基质中。通过该过程产生尺寸规则的油颗粒，其直径的变化系数（S.D./平均）为16％。这种变化大于使用先前的微通道阵列在先前的试验中获得的变化（最小1-2％），这似乎是由于锯切造成的微切屑导致了通道末端不规则。作为相对于先前的优点，本发明的微通道阵列允许在过程中容易收集产生的油颗粒，并且还容易改变悬浮流体的组成。因此，表明该衬底的侧面可用于用传统的或手动的工具或方法在该衬底外部的微结构化在主衬底表面中的结构或装置，该结构或装置可用玻璃板覆盖。 ！16

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