Intuitive, image-based cell sorting using optofluidic cell sorting
Intuitive, image-based cell sorting using optofluidic cell sorting
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DOI：
10.1021/ac071366y
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发表时间：
2007-12-15
影响因子：
7.4
通讯作者：
Voldman, J.
中科院分区：
文献类型：
--
作者：
Kovac, J. R.;Voldman, J.
我们提出了一种微流体细胞分选装置，它增强了显微镜的能力，可以进行简单的基于图像的细胞分选。朱鹭的组合使基于时空荧光或细胞形态的直观、复杂的表型分类成为可能。微流控装置包含一个微孔阵列，可以通过沉降被动加载哺乳动物细胞，随后可以用显微镜检查。检查后，我们利用聚焦红外激光的散射力将感兴趣的细胞从井中悬浮到流场中进行收集。首先，我们展示了基于全细胞荧光的基于图像的分选，这可以实现基于时间全细胞荧光行为的分选。其次，我们展示了基于荧光定位（核荧光与全细胞荧光）的基于图像的分类，突出了我们的方法基于图像亚细胞事件（如局部蛋白表达或易位事件）分类的能力。我们的分选后纯度高达89%，目标细胞的富集度高达155倍。光学操作文献和直接细胞活力测定表明，细胞在使用我们的技术后仍然具有活力。该架构具有高度可扩展性，支持超过10,000个可单独寻址的陷阱站点。我们的方法能够基于亚细胞时空信息对重要种群进行排序，这对于现有的广泛分类技术来说是困难或不可能的。
We present a microfluidic cell-sorting device which augments microscopy with the capability to perform facile image-based cell sorting. Ibis combination enables intuitive, complex phenotype sorting based on spatio-temporal fluorescence or cell morphology. The microfluidic device contains a microwell array that can be passively loaded with mammalian cells via sedimentation and can be subsequently inspected with microscopy. After inspection, we use the scattering force from a focused infrared laser to levitate cells of interest from their wells into a flow field for collection. First, we demonstrate image-based sorting predicated on whole-cell fluorescence, which could enable sorting based on temporal whole-cell fluorescence behavior. Second, we demonstrate image-based sorting predicated on fluorescence localization (nuclear vs whole-cell fluorescence), highlighting the capability of our approach to sort based on imaged subcellular events' such as localized protein expression or translocation events. We achieve postsort purities up to 89% and up to 155-fold enrichment of target cells. Optical manipulation literature and a direct cell viability assay suggest that cells remain viable after using our technique. The architecture is highly scalable and supports over 10 000 individually addressable trap sites. Our approach enables sorting of significant populations based on subcellular spatio-temporal information, which is difficult or impossible with existing widespread sorting technologies.