Localized substance delivery to single cell and 4D imaging of its uptake using a flow channel with a lateral aperture
Localized substance delivery to single cell and 4D imaging of its uptake using a flow channel with a lateral aperture
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DOI：
10.1007/s10404-011-0885-3
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发表时间：
2012-01-01
影响因子：
2.8
通讯作者：
Kotera, Hidetoshi
中科院分区：
文献类型：
--
作者：
Terao, Kyohei;Okonogi, Atsuhito;Kotera, Hidetoshi
我们开发了一种新型的微流体装置，用于当细胞表面部分暴露于外部刺激时，对细胞内事件进行空间和时间分辨（4D）可视化。该装置使用光刻胶SU-8的3D旋转倾斜UV光刻技术制造，由包含细胞的腔室和由具有比细胞小的横向微米孔径的薄垂直壁分隔的流动通道组成。首先使用注射泵从流动通道中吸出细胞，将细胞固定在孔上，然后将化学刺激物馈送到通道中，以便仅由孔包围的细胞表面受到刺激而不泄漏到细胞表面的其他部分。随后的横向信号传播在细胞内可以可视化使用高速荧光共聚焦显微镜。作为该装置的实验演示，以4D分辨率显示2-NBDG（荧光葡萄糖类似物）摄入小鼠胰岛素瘤细胞MIN6m9。
We have developed a novel microfluidic device for space- and time-resolved (4D) visualization of intracellular events when a cell surface is partially exposed to external stimuli. The device, fabricated using 3D rotational inclined UV lithography of photoresist SU-8, consists of a cell-containing chamber and a flow channel separated by a thin vertical wall having a lateral micrometer aperture smaller than a cell. A cell is first immobilized on the aperture by suction from the flow channel using a syringe pump, and a chemical stimulant is then fed to the channel so that only the cell surface bounded by the aperture is subjected to the stimulus without leakage to other part of the cell surface. The subsequent lateral signal propagation inside the cell can be visualized using high-speed fluorescence confocal microscopy. As an experimental demonstration of the device, 2-NBDG (fluorescence glucose analog) intake into a mouse insulinoma cell, MIN6m9, was visualized in 4D resolution.