Live cell lithography: Using optical tweezers to create synthetic tissue
Live cell lithography: Using optical tweezers to create synthetic tissue
复制标题
DOI：
10.1039/b807987k
复制
发表时间：
2008-01-01
期刊：
影响因子：
6.1
通讯作者：
Timp, Gregory
中科院分区：
文献类型：
--
作者：
Mirsaidov, Utkur;Scrimgeour, Jan;Timp, Gregory
我们展示了一种创建合成组织的新方法，该方法有可能以亚微米精度捕获多细胞生物体的三维 (3D) 复杂性。我们在微流体网络中使用多个层流流体流，将细胞传送到组装区域，在该区域中使用多个分时光镊将它们组织成复杂的阵列。然后将细胞封装在模仿细胞外基质的 30 μm x 30 μm x 45 μm 体积的光聚合水凝胶中。为了在不损失可行性的情况下扩展阵列的大小、形状和选区，我们然后走到相邻位置，同时保持与参考阵列的配准，并重复该过程。使用这种分步重复方法，我们形成了一个异质的大肠杆菌阵列，该阵列经过基因工程改造，具有与荧光报告基因功能相连的紫胶开关。然后，我们通过微流体网络使用配体诱导阵列，并跟踪荧光的时空发展以评估活力和代谢活性。
We demonstrate a new method for creating synthetic tissue that has the potential to capture the three-dimensional (3D) complexity of a multi-cellular organism with submicron precision. Using multiple laminar fluid flows in a microfluidic network, we convey cells to an assembly area where multiple, time-shared optical tweezers are used to organize them into a complex array. The cells are then encapsulated in a 30 mu m x 30 mu m x 45 mu m volume of photopolymerizable hydrogel that mimicks an extra-cellular matrix. To extend the size, shape and constituency of the array without loss of viability, we then step to an adjacent location while maintaining registration with the reference array, and repeat the process. Using this step-and-repeat method, we formed a heterogeneous array of E. coli genetically engineered with a lac switch that is functionally linked to fluorescence reporters. We then induced the array using ligands through a microfluidic network and followed the space-time development of the fluorescence to evaluate viability and metabolic activity.