![]() This dissertation aims to develop a droplet-based artificial cell system using cell-free extracts of Xenopus laevis eggs and understand mitotic oscillations with the proposed system. ![]() We demonstrate the applicability of the integrated device to synthetic biology through sequentially fusing SUVs with freshly prepared GUVs and separating the fused GUVs from extraneous SUVs and oil droplets at the same time. In addition, by incorporating the purification module with a microfluidic-based GUV-formation method, octanol-assisted liposome assembly (OLA), we established an integrated production-purification microfluidic unit that sequentially produces, manipulates, and purifies GUVs. Using our purification approach, we showed that giant vesicles can be separated from various residues, that range in size and chemical composition, with a very high efficiency (e=0.99), based on size and deformability of the filtered objects. We developed a microfluidic platform capable of purifying GUVs through stream bifurcation, where a stream of vesicles suspension is partitioned into three fractions - purified GUVs, residual components, and a washing solution. Therefore, with the rapidly growing field of synthetic biology, there is an emergent demand for techniques that can continuously purify cell-like vesicles from diverse residues, while GUVs are being simultaneously synthesized and manipulated. However, the presence of residual components and by-products, generated during vesicles preparation and manipulation, severely limits the utility of GUVs in applications like synthetic cells. ![]() This new device minimizes the sample consumption and could therefore be applied for studies using precious samples.Ĭell-sized vesicles like giant unilamellar vesicles (GUVs) are established as a promising biomimetic model for studying cellular phenomena in isolation. Importantly, the devices were fabricated using a classical soft-lithography method, as opposed to additive manufacturing often used to generate complex mixing structures. The most promising design was then visualized experimentally, showing results that are consistent with the outcomes of the simulations.
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