Photostick: a method for selective isolation of target cells from culture.
Photostick: a method for selective isolation of target cells from culture.
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DOI：
10.1039/c4sc03676j
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发表时间：
2015-03
期刊：
影响因子：
8.4
通讯作者：
Cohen AE
中科院分区：
文献类型：
--
作者：
Chien MP;Werley CA;Farhi SL;Cohen AE
一种新的方法能够通过与培养皿的光化学交联来光学选择培养中的一个或多个细胞。当选择标准复杂时，例如动态响应、形态学特征或多个参数的组合，从异质池中分选靶细胞在技术上是困难的。目前，哺乳动物细胞选择通常通过静态荧光（例如荧光激活细胞分选仪）、通过存活（例如抗生素抗性）或通过连续操作（流式细胞术、激光捕获显微切割）来进行。在这里，我们提出了一个简单的协议，用于选择细胞的基础上，可以通过视频显微镜和图像处理确定的任何静态或动态的属性。“photostick”技术使用细胞不渗透的光化学交联剂和基于数字阵列的图案化照明来将选定的细胞固定在培养皿上。其他细胞用温和的蛋白酶处理洗掉。交联剂还用荧光染料和生物素标记所选择的细胞，用于以后的鉴定。光敏棒方案保留细胞活力，允许对所选细胞进行遗传分析，并且可以用复杂的功能选择标准如神经元放电模式进行。
A new method enables optical selection of one or more cells in culture via photochemical crosslinking to the culture dish. Sorting of target cells from a heterogeneous pool is technically difficult when the selection criterion is complex, e.g. a dynamic response, a morphological feature, or a combination of multiple parameters. At present, mammalian cell selections are typically performed either via static fluorescence (e.g. fluorescence activated cell sorter), via survival (e.g. antibiotic resistance), or via serial operations (flow cytometry, laser capture microdissection). Here we present a simple protocol for selecting cells based on any static or dynamic property that can be identified by video microscopy and image processing. The “photostick” technique uses a cell-impermeant photochemical crosslinker and digital micromirror array-based patterned illumination to immobilize selected cells on the culture dish. Other cells are washed away with mild protease treatment. The crosslinker also labels the selected cells with a fluorescent dye and a biotin for later identification. The photostick protocol preserves cell viability, permits genetic profiling of selected cells, and can be performed with complex functional selection criteria such as neuronal firing patterns.