Production of novel proteins using the micro system that permits sorting and functional evaluation at single molecular level

我们研制了一种以温敏水凝胶为微阀的微型荧光激活生物分子分选器。该分选机由一个入口和两个出口的Y形通道的微流控芯片组成。将含有荧光标记细胞和热敏聚合物的生物标本引入Y形微通道。热敏聚合物受热后，由溶胶状态变为凝胶状态。通过定点红外激光照射，局部诱导溶胶-凝胶转变，以堵塞微通道中的一个并收集细胞。作为该分选器的一个应用，表达GFP的E.coll.分离荧光抗体标记的CD8阳性淋巴细胞和TOTO-1标记的DNA分子。该分选机分选时间为120ms，回收率为90%。

We have developed a micro fabricated fluorescence-activated bio-molecular sorter using thermosensitive hydrogel as a microvalve. The sorter was composed of a microfluidic chip with Y-shaped channels with one inlet and two outlets. Biological specimen containing fluorescently labeled cells and thermosensitive polymer was introduced into a Y-shaped microchannel. Thermosensitive polymer changed the state from sol to gel when it was heated. The sol-gel transition was locally induced by site directed infrared laser irradiation in order to plug the one of the microchannel and collect the cells. As an application of this sorter, GFP-expressing E. coll. CD8-expressing lymphocytes labeled with fluorescence antibody, and DNA molecules labeled with TOTO-1 were separated. The sorting time of 120 ms and recovery ratio of 90 % was achieved by this sorter.