Optical painting and fluorescence activated sorting of single adherent cells labelled with photoswitchable Pdots.
Optical painting and fluorescence activated sorting of single adherent cells labelled with photoswitchable Pdots.
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DOI：
10.1038/ncomms11468
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发表时间：
2016-04-27
影响因子：
16.6
通讯作者：
Chiu DT
中科院分区：
文献类型：
--
作者：
Kuo CT;Thompson AM;Gallina ME;Ye F;Johnson ES;Sun W;Zhao M;Yu J;Wu IC;Fujimoto B;DuFort CC;Carlson MA;Hingorani SR;Paguirigan AL;Radich JP;Chiu DT
在许多生物医学和临床应用中，期望从混合群体中有效地选择和分离感兴趣的单个细胞。在这里，我们展示了使用光开关半导体聚合物点（Pdots）作为光学“绘画”工具的概念，这使得能够在显微镜下根据其荧光及其空间和形态特征选择某些粘附细胞。我们首先开发了一种Pdot，它可以在亮（ON）和暗（OFF）状态之间可逆地切换，在紫外光或红光照射下具有150倍的对比度。使用聚焦的633 nm激光束作为“画笔”，光可转换Pdot作为“油漆”，我们通过打开荧光选择和“油漆”单个Pdot标记的贴壁细胞，然后进行分类和回收光学标记的细胞（回收率为90%，纯度接近100%），然后进行遗传分析。 从混合群体中分离单个细胞对于许多生物医学应用是期望的。在这里，作者使用可光切换的Pdot来实现“光学绘画”，其中感兴趣的细胞基于其视觉特征进行标记，然后可以通过荧光激活细胞分选进行分离。
The efficient selection and isolation of individual cells of interest from a mixed population is desired in many biomedical and clinical applications. Here we show the concept of using photoswitchable semiconducting polymer dots (Pdots) as an optical ‘painting' tool, which enables the selection of certain adherent cells based on their fluorescence, and their spatial and morphological features, under a microscope. We first develop a Pdot that can switch between the bright (ON) and dark (OFF) states reversibly with a 150-fold contrast ratio on irradiation with ultraviolet or red light. With a focused 633-nm laser beam that acts as a ‘paintbrush' and the photoswitchable Pdots as the ‘paint', we select and ‘paint' individual Pdot-labelled adherent cells by turning on their fluorescence, then proceed to sort and recover the optically marked cells (with 90% recovery and near 100% purity), followed by genetic analysis. Isolation of individual cells from mixed populations is desirable for many biomedical applications. Here the authors use photoswitchable Pdots to allow 'optical painting', where cells of interest are marked based on their visual characteristics, and can then be isolated by fluorescence activated cell sorting.