Department of Electrical Engineering and Computer Science, Massachusetts Institute of Technology, Cambridge, Massachusetts 02139, USA.
Nat Commun. 2011;2:271. doi: 10.1038/ncomms1266.
High-resolution in vivo time-lapse assays require repeated immobilization and imaging of whole animals. Here we report a technology for screening Caenorhabditis elegans at cellular resolution over its entire lifespan inside standard multiwell plates using repeated immobilization, imaging and optical manipulation. Our system does not use any fluidic or mechanical components, and can operate for tens of thousands of cycles without failure. It is also compatible with industrial high-throughput screening platforms and robotics, and it allows both chemical, and forward and reverse genetic screens. We used this technology to perform subcellular-resolution femtosecond laser microsurgery of single neurons in vivo, and to image the subsequent regeneration dynamics at subcellular resolution. Our single-neuron in vivo time-lapse analysis shows that neurite regrowth occurring over short time windows is significantly greater than that predicted by ensemble averaging over many animals.
高分辨率的活体延时分析需要对整个动物进行重复的固定和成像。在这里，我们报告了一种在标准多孔板中以细胞分辨率筛选秀丽隐杆线虫的技术，该技术可通过重复固定、成像和光学操作来实现。我们的系统不使用任何流体或机械部件，并且可以在不发生故障的情况下运行数万次。它还与工业高通量筛选平台和机器人兼容，并且允许进行化学筛选以及正向和反向遗传筛选。我们使用这项技术对单个神经元进行了亚细胞分辨率的飞秒激光显微手术，并以亚细胞分辨率对随后的再生动力学进行了成像。我们的单细胞活体延时分析表明，在短时间窗口内发生的神经突再生明显大于通过对许多动物进行平均预测的值。
