Shiomi Hideaki, Tsuda Soichiro, Suzuki Hiroaki, Yomo Tetsuya
Graduate School of Information Science and Technology, Osaka University, Osaka, Japan.
School of Chemistry, University of Glasgow, Glasgow, United Kingdom; ERATO, JST, Tokyo, Japan.
PLoS One. 2014 Jul 3;9(7):e101820. doi: 10.1371/journal.pone.0101820. eCollection 2014.
This paper describes the utilization of giant unilamellar vesicles (GUVs) as a platform for handling chemical and biochemical reagents. GUVs with diameters of 5 to 10 µm and containing chemical/biochemical reagents together with inert polymers were fused with electric pulses (electrofusion). After reagent mixing, the fused GUVs spontaneously deformed to a budding shape, separating the mixed solution into sub-volumes. We utilized a microfluidic channel and optical tweezers to select GUVs of interest, bring them into contact, and fuse them together to mix and aliquot the reaction product. We also show that, by lowering the ambient temperature close to the phase transition temperature Tm of the lipid used, daughter GUVs completely detached (fission). This process performs all the liquid-handing features used in bench-top biochemistry using the GUV, which could be advantageous for the membrane-related biochemical assays.
本文描述了利用巨型单层囊泡(GUVs)作为处理化学和生化试剂的平台。直径为5至10微米、含有化学/生化试剂以及惰性聚合物的GUVs通过电脉冲(电融合)进行融合。试剂混合后,融合的GUVs自发变形为出芽形状,将混合溶液分离成子体积。我们利用微流控通道和光镊来选择感兴趣的GUVs,使其接触并融合在一起,以混合和分装反应产物。我们还表明,通过将环境温度降低至接近所用脂质的相变温度Tm,子代GUVs会完全分离(裂变)。该过程利用GUV实现了台式生物化学中使用的所有液体处理功能,这对于与膜相关的生化分析可能具有优势。