Intravital microscopy of biosensor activities and intrinsic metabolic states
Intravital microscopy of biosensor activities and intrinsic metabolic states
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DOI：
10.1016/j.ymeth.2017.04.017
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发表时间：
2017-09-01
期刊：
影响因子：
4.8
通讯作者：
Day, Richard N.
中科院分区：
文献类型：
--
作者：
Winfree, Seth;Hato, Takashi;Day, Richard N.
活体显微镜（IVM）是一种成像工具，能够检测活体动物组织中发生的亚细胞信号或代谢事件。然而，在高度散射的生物组织中成像是具有挑战性的，因为在增加的深度处获取的图像中的信号衰减。与深度有关的信号衰减是IVM的主要障碍，限制了可以获得重要数据的深度。因此，通过IVM进行定量测量需要使用内部校准的方法，或者使用完全不同的方法来评估信号。在这里，我们描述了如何比率成像的基因编码的生物传感器探针可以用来定量测量细胞信号通路的活性变化。然后，我们描述了荧光寿命成像可以用于活体动物内细胞代谢状态的无标记测量。(C)2017由Elsevier Inc.出版
Intravital microscopy (IVM) is an imaging tool that is capable of detecting subcellular signaling or metabolic events as they occur in tissues in the living animal. Imaging in highly scattering biological tissues, however, is challenging because of the attenuation of signal in images acquired at increasing depths. Depth-dependent signal attenuation is the major impediment to IVM, limiting the depth from which significant data can be obtained. Therefore, making quantitative measurements by IVM requires methods that use internal calibration, or alternatively, a completely different way of evaluating the signals. Here, we describe how ratiometric imaging of genetically encoded biosensor probes can be used to make quantitative measurements of changes in the activity of cell signaling pathways. Then, we describe how fluorescence lifetime imaging can be used for label-free measurements of the metabolic states of cells within the living animal. (C) 2017 Published by Elsevier Inc.