Selective encapsulation of single cells and subcellular organelles into picoliter- and femtoliter-volume droplets
Selective encapsulation of single cells and subcellular organelles into picoliter- and femtoliter-volume droplets
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DOI：
10.1021/ac0480850
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发表时间：
2005-03-15
影响因子：
7.4
通讯作者：
Chiu, DT
中科院分区：
文献类型：
--
作者：
He, MY;Edgar, JS;Chiu, DT
Ibis论文描述了一种方法，其结合了光学捕获和基于微流体的液滴产生，用于选择性地和可控地将单个靶细胞或亚细胞结构（例如微囊体）包封到由不混溶相包围的皮升或飞升体积的水性液滴中。一旦选定的细胞或细胞器被包裹在液滴内，它就被稳定地限制在液滴中并且不能被移除。我们在液滴中证明了单细胞的快速激光光解，其基本上“冻结”了细胞在光解时刻所处的状态，并将裂解物限制在液滴的小体积内。使用荧光素二-β-D-吡喃半乳糖苷，这是一种荧光底物的细胞内酶β-半乳糖苷酶，我们还测定了这种酶的活性从一个单一的细胞后，激光诱导的细胞裂解。Ibis捕获单个选定细胞或亚细胞器的能力应该为进行单细胞研究和单细胞器测量开辟新的可能性。
Ibis paper describes a method, which combines optical trapping and microfluidic-based droplet generation, for selectively,and controllably encapsulating a single target cell or subcellular structure, such as a mitochondrion, into a picoliter- or femtoliter-volume aqueous droplet that is surrounded by an immiscible phase. Once the selected cell or organelle is encased within the droplet, it is stably confined in the droplet and cannot be removed. We demonstrate in droplet the rapid laser photolysis of the single cell, which essentially "freezes" the state that the cell was in at the moment of photolysis and confines the lysate within the small volume of the droplet. Using fluorescein di-beta-D-galactopyranoside, which is a fluorogenic substrate for the intracellular enzyme beta-galactosidase, we also assayed the activity of this enzyme from a single cell following the laser-induced lysis of the cell. Ibis ability to entrap individual selected cells or subcellular organelles should open new possibilities for carrying out single-cell studies and single-organelle measurements.