Two-photon probes for in vivo multicolor microscopy of the structure and signals of brain cells.
Two-photon probes for in vivo multicolor microscopy of the structure and signals of brain cells.
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DOI：
10.1007/s00429-018-1678-1
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发表时间：
2018-09
影响因子：
3.1
通讯作者：
Fiole D
中科院分区：
文献类型：
--
作者：
Ricard C;Arroyo ED;He CX;Portera-Cailliau C;Lepousez G;Canepari M;Fiole D
由于所用激发波长的高穿透性和低光毒性，可以通过双光子显微镜在微观尺度上对活体实验室动物的大脑进行成像。然而，知识的双光子光谱特性的无数荧光探针一般是稀缺的，对于许多人来说，不存在。此外，在已发表的报告中使用不同的测量单位进一步阻碍了综合成像实验的设计。在这篇综述中，我们编译和均匀化的280荧光探针的双光子光谱特性。我们提供了实际数据，包括最佳双光子激发波长，双光子作用截面或分子亮度的峰值，和发射范围。除了这些荧光团的光谱描述之外，我们还讨论了它们与生物靶点的结合。这种特异性允许在体内成像的细胞，其过程，甚至细胞器和其他亚细胞结构在大脑中。除了监测内源性细胞代谢的探针外，健康和患病大脑的研究还受益于某些探针与病理学特异性特征的特异性结合，从淀粉样蛋白-β斑块到某些抗生素的自发荧光。一个特别的重点放在功能性的体内成像使用双光子探针，感测特定的离子或膜电位，并可能与光遗传致动器相结合。与它们的用途密切相关，我们根据目标检查这些荧光探针的活体内递送的不同途径。最后，我们讨论了不同的方法，策略和先决条件，在活体实验室动物的大脑中进行双光子双光子成像实验。
Imaging the brain of living laboratory animals at a microscopic scale can be achieved by two-photon microscopy thanks to the high penetrability and low phototoxicity of the excitation wavelengths used. However, knowledge of the two-photon spectral properties of the myriad fluorescent probes is generally scarce and, for many, non-existent. Additionally, the use of different measurement units in published reports further hinders the design of a comprehensive imaging experiment. In this review, we compile and homogenize the two-photon spectral properties of 280 fluorescent probes. We provide practical data, including the wavelengths for optimal two-photon excitation, the peak values of two-photon action cross-section or molecular brightness, and the emission ranges. Beyond the spectroscopic description of these fluorophores, we discuss their binding to biological targets. This specificity allows in vivo imaging of cells, their processes, and even organelles and other subcellular structures in the brain. In addition to probes that monitor endogenous cell metabolism, studies of healthy and diseased brain benefit from the specific binding of certain probes to pathology-specific features, ranging from amyloid-β plaques to the autofluorescence of certain antibiotics. A special focus is placed on functional in vivo imaging using two-photon probes that sense specific ions or membrane potential, and that may be combined with optogenetic actuators. Being closely linked to their use, we examine the different routes of intravital delivery of these fluorescent probes according to the target. Finally, we discuss different approaches, strategies, and prerequisites for two-photon multicolor experiments in the brains of living laboratory animals.
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影响因子：
3.4
作者：
Ando, Ryoko;Flors, Cristina;Miyawaki, Atsushi
通讯作者：
Miyawaki, Atsushi
影响因子：
3.4
作者：
Brinks, Daan;Klein, Aaron J.;Cohen, Adam E.
通讯作者：
Cohen, Adam E.
影响因子：
48
作者：
Ai, Hui-wang;Hazelwood, Kristin L.;Campbell, Robert E.
通讯作者：
Campbell, Robert E.
影响因子：
3.5
作者：
Anderson VL;Webb WW
通讯作者：
Webb WW
影响因子：
5.5
作者：
Brain, KL;Bennett, MR
通讯作者：
Bennett, MR