Automated on-chip rapid microscopy, phenotyping and sorting of C. elegans
Automated on-chip rapid microscopy, phenotyping and sorting of C. elegans
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DOI：
10.1038/nmeth.1227
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发表时间：
2008-07-01
期刊：
影响因子：
48
通讯作者：
Lu, Hang
中科院分区：
文献类型：
--
作者：
Chung, Kwanghun;Crane, Matthew M.;Lu, Hang
显微镜、表型分析和视觉筛选经常与遗传学结合应用于模式生物。虽然被广泛使用，但这些用于多细胞生物体的技术大多仍然是手动和低通量的。在这里，我们报告的样品处理，高分辨率显微镜，表型和分类秀丽隐杆线虫的完全自动化。工程微流体系统，加上定制的软件，使高通量，高分辨率显微镜和分选没有人为干预，并可以与任何显微镜设置相结合。该微芯片能够进行强大的局部温度控制，自我调节样品加载和自动样品定位，而集成软件则根据形态和强度特征对蠕虫进行成像和分类。我们证明了基于细胞和亚细胞表型进行灵敏和定量筛选的能力，每轮准确率超过95%，每小时数百只蠕虫的速度。筛选时间可以减少几个数量级;此外，筛选是完全自动化的。
Microscopy, phenotyping and visual screens are frequently applied to model organisms in combination with genetics. Although widely used, these techniques for multicellular organisms have mostly remained manual and low-throughput. Here we report the complete automation of sample handling, high-resolution microscopy, phenotyping and sorting of Caenorhabditis elegans. The engineered microfluidic system, coupled with customized software, has enabled high-throughput, high-resolution microscopy and sorting with no human intervention and may be combined with any microscopy setup. The microchip is capable of robust local temperature control, self-regulated sample-loading and automatic sample-positioning, while the integrated software performs imaging and classification of worms based on morphological and intensity features. We demonstrate the ability to perform sensitive and quantitative screens based on cellular and subcellular phenotypes with over 95% accuracy per round and a rate of several hundred worms per hour. Screening time can be reduced by orders of magnitude; moreover, screening is completely automated.