Live Cell Imaging Using Photoswitchable Diarylethene-Doped Fluorescent Polymer Dots
Live Cell Imaging Using Photoswitchable Diarylethene-Doped Fluorescent Polymer Dots
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DOI：
10.1002/asia.201701038
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发表时间：
2017-10-18
影响因子：
4.1
通讯作者：
Majima, Tetsuro
中科院分区：
文献类型：
--
作者：
Osakada, Yasuko;Fukaminato, Tuyoshi;Majima, Tetsuro
近年来，利用纳米材料的荧光光开关已成为生物靶标成像的一种很有前途的方法。然而，尽管在过去十年中进行了密集的研究工作，但使用可光开关纳米颗粒实时应用于生物靶标的实用显微镜仍然具有挑战性。为了解决这个问题，我们开发了活的巨噬细胞成像和单粒子成像方法，在氙灯照射下使用可光开关的荧光二芳基乙烯掺杂聚合物纳米颗粒(P-Dot)。在氙灯和绿色荧光蛋白滤光片的可见光照射下，我们用掺杂了二芳基乙烯甲氧基的P-点建立了34倍长的“关态”。为了证明掺杂P-点成像的实用性，我们对巨噬细胞中的溶酶体进行了成像，观察到从关闭状态到开启状态的荧光恢复速度慢了11倍，这表明了它们用于细胞成像的潜力。
Fluorescence photoswitching using nanomaterials has recently emerged as a promising approach for the imaging of biological targets. However, despite intensive research efforts during the last decade, practical microscopy of biological targets using photoswitchable nanoparticles in real time remains challenging. To address this problem, we have developed live macrophage cell imaging and single particle imaging methods, using photoswitchable fluorescent diarylethene-doped polymer nanoparticles (P-dots) under Xe lamp irradiation. We established a 34-times prolonged "off-state", using P-dots doped with a diarylethene-containing methoxy substituent, upon visible-light irradiation using a Xe lamp and a green fluorescent protein filter cube. To demonstrate the practicality of doped P-dots imaging, we imaged lysosomes in macrophage cells, and observed 11-times slower recovery of the fluorescence from the off-state to the "on-state", indicating their potential for cellular imaging.