On-chip cell sorting system using laser-induced heating of a thermoreversible gelation polymer to control flow
On-chip cell sorting system using laser-induced heating of a thermoreversible gelation polymer to control flow
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DOI：
10.1021/ac0511041
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发表时间：
2006-02-01
影响因子：
7.4
通讯作者：
Funatsu, T
中科院分区：
文献类型：
--
作者：
Shirasaki, Y;Tanaka, J;Funatsu, T
我们开发了一种微制造荧光激活细胞分选系统，使用热可逆凝胶化聚合物（TGP）作为开关阀。该玻璃分选芯片具有具有一个入口和两个出口的y形微通道。将含有荧光标记细胞的生物标本与含有热可逆溶胶-凝胶聚合物的溶液混合。然后将混合溶液通过入口引入分选机芯片。利用红外激光定点照射堵塞其中一个出口，局部诱导溶胶-凝胶转化。用灵敏荧光显微镜检测荧光标记的靶细胞。在没有荧光信号的情况下，通过激光照射TGP堵塞收集通道，将标本引导至废通道。一旦检测到来自目标细胞的荧光信号，激光束就被用来堵塞废物通道，使荧光细胞进入收集池。溶胶-凝胶转化的响应时间为3 ms，流动切换时间为120 ms。利用该系统，我们演示了荧光微球和表达荧光蛋白的大肠杆菌细胞的分选。从分选系统中提取后发现这些细胞是有活力的，表明细胞没有受到损伤。
We have developed a microfabricated fluorescence-activated cell sorter system using a thermoreversible gelation polymer (TGP) as a switching valve. The glass sorter chip has Y-shaped microchannels with one inlet and two outlets. A biological specimen containing fluorescently labeled cells is mixed with a solution containing a thermoreversible sol-gel polymer. The mixed solution is then introduced into the sorter chip through the inlet. The sol-gel transformation was locally induced by site-directed infrared laser irradiation to plug one of the outlets. The fluorescently labeled target cells were detected with sensitive fluorescence microscopy. In the absence of a fluorescence signal, the collection channel is plugged through laser irradiation of the TGP and the specimens are directed to the waste channel. Upon detection of a fluorescence signal from the target cells, the laser beam is then used to plug the waste channel, allowing the fluorescent cells to be channeled into the collection reservoir. The response time of the sol-gel transformation was 3 ms, and a flow switching time of 120 ms was achieved. Using this system, we have demonstrated the sorting of fluorescent microspheres and Escherichia coli cells expressing fluorescent proteins. These cells were found to be viable after extraction from the sorting system, indicating no damage to the cells.