用在细胞和组织中的双光子化探针检测酸酶活动的有机体特异性检测
1Department of Chemistry, National University of Singapore, Singapore 117543.
Journal of the American Chemical Society
|June 28, 2012
概括
研究人员开发了一种新型的小分子记者系统,用于双光子光显微镜 (TPFM),以图像活细胞和Drosophila大脑中的酶活性. 该系统为生物研究提供了增强的成像深度和特异性.
科学领域:
- 生物成像是一种生物成像.
- 分子成像学分子成像学
- 生物化学 生物化学
背景情况:
- 与传统方法相比,双光子光显微镜 (TPFM) 提供了更高的深度透率和更少的光损伤.
- 小分子探针在生物成像中比光蛋白 (FP) 具有优势,原因是光物理特性和细胞透性.
研究的目的:
- 开发和描述一种新的化,小分子记者系统,用于成像内源性酸酶活动.
- 用TPFM证明这种系统对活哺乳动物细胞和虫大脑的实用性.
主要方法:
- 研究了Y2/Y1记者系统的1和2光子激发光物理性质.
- 将记者系统与细胞透 (CPPs) 结合起来,用于向成像.
- 利用TPFM对活细胞和Drosophila大脑中的内源酸酶活动进行成像.
主要成果:
- Y2/Y1系统显示了适合TPFM的Turn-ON光特性.
- 实现了高空间和时间分辨率的酸酶活动的有机细胞和瘤细胞特异成像.
- 在Drosophila大脑中成功成像了内源性酸酶活动,深度超过100微米.
结论:
- 开发的记者系统是第一个从具有有利的两光子特性的中心对称染料中设计的酶报告TPFM系统.
- 该系统有效地克服了小分子探针的扩散限制.
- 这种新的TPFM系统能够在体内对酶活性进行深层组织成像.
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