Three dimensional live cell lithography
Three dimensional live cell lithography
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DOI：
10.1364/oe.21.010269
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发表时间：
2013-04-22
期刊：
影响因子：
3.8
通讯作者：
McLeod, Robert R.
中科院分区：
文献类型：
--
作者：
Linnenberger, Anna;Bodine, Martha I.;McLeod, Robert R.
我们研究了全息光学捕获与步进重复无掩模投影立体光刻相结合，以精细控制三维微结构水凝胶内活细胞的三维位置。选择C2C12成肌细胞作为演示平台，因为成肌细胞向多核肌管的发育需要成肌细胞的线性排列。C2C12细胞被放置在1064 nm处具有多个光阱的单体溶液中，然后通过投射一个512 × 512空间光调制器在405 nm处进行单体光聚合封装。采用LAP盐光引发剂可实现405 nm的高灵敏度和1064 nm的完全不敏感。这些波长，加上白光LED的亮场成像，可以通过一个油浸物镜同时聚焦。通过x和y步进重复过程实现了大的横向尺寸的图案凝胶/细胞结构。通过多层立体光刻实现了大厚度，允许制造具有微米级结构和毫米尺寸的精确排列的3D活细胞支架。在诱捕和包封过程后，细胞仍能保持活力。(C) 2013年美国光学学会
We investigate holographic optical trapping combined with step-and-repeat maskless projection stereolithography for fine control of 3D position of living cells within a 3D microstructured hydrogel. C2C12 myoblast cells were chosen as a demonstration platform since their development into multinucleated myotubes requires linear arrangements of myoblasts. C2C12 cells are positioned in the monomer solution with multiple optical traps at 1064 nm and then encapsulated by photopolymerization of monomer via projection of a 512x512 spatial light modulator illuminated at 405 nm. High 405 nm sensitivity and complete insensitivity to 1064 nm was enabled by a lithium acylphosphinate (LAP) salt photoinitiator. These wavelengths, in addition to brightfield imaging with a white light LED, could be simultaneously focused by a single oil immersion objective. Large lateral dimensions of the patterned gel/cell structure are achieved by x and y step-and-repeat process. Large thickness is achieved through multi-layer stereolithography, allowing fabrication of precisely-arranged 3D live cell scaffolds with micron-scale structure and millimeter dimensions. Cells are shown to retain viability after the trapping and encapsulation procedure. (C) 2013 Optical Society of America