Ontario Cancer Institute and Techna Institute, University Health Network, Toronto, Ontario M5G1L7, Canada, Department of Medical Biophysics, University of Toronto, Ontario, M5G1L7, Canada.
Small. 2014 Mar 26;10(6):1184-93. doi: 10.1002/smll.201302424. Epub 2013 Dec 21.
Efforts to develop self-contained microreactors and artificial cells have been limited by difficulty in generating membranes that can be robustly and repeatedly manipulated to load and release cargo from phospholipid compartments. Here we describe a purely optical method to form pores in a membrane generated from porphyrin-phospholipid conjugates electro-assembled into microscale giant porphyrin vesicles and manipulated using confocal microscopy. The pores in the membrane resealed within a minute allowing for repeated pore formation with precise spatial and temporal control and optical gating to allow selective diffusion of biomolecules across the membrane. Temporal control of pore formation was illustrated by performing sequential DNA hybridization reactions. A biotin-avidin based strategy was developed to selectively attach enzymes to the interior of the vesicle, demonstrating spatial control and the potential of giant porphyrin vesicles as versatile microreactors.
开发独立式微反应器和人工细胞的努力受到限制，因为难以生成能够稳定且可重复地操纵的膜，以将货物从磷脂隔室中装载和释放。在这里，我们描述了一种纯光学方法，用于在由 porphyrin-phospholipid 缀合物电组装成微尺度 giant porphyrin 囊泡的膜中形成孔，并使用共聚焦显微镜进行操纵。膜中的孔在一分钟内重新密封，允许进行重复的孔形成，具有精确的空间和时间控制以及光学门控，以允许生物分子选择性地穿过膜扩散。通过进行顺序 DNA 杂交反应来说明孔形成的时间控制。开发了一种基于生物素-亲和素的策略，将酶选择性地连接到囊泡的内部，证明了空间控制和 giant porphyrin 囊泡作为多功能微反应器的潜力。
