Live-Cell Bioorthogonal Chemical Imaging: Stimulated Raman Scattering Microscopy of Vibrational Probes.
Live-Cell Bioorthogonal Chemical Imaging: Stimulated Raman Scattering Microscopy of Vibrational Probes.
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DOI：
10.1021/acs.accounts.6b00210
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发表时间：
2016-08-16
影响因子：
18.3
通讯作者：
Min W
中科院分区：
文献类型：
--
作者：
Wei L;Hu F;Chen Z;Shen Y;Zhang L;Min W
光学显微镜的创新极大地改变了研究人员以亚细胞分辨率研究生物系统的方式。特别是，荧光显微镜与荧光探针的扩大选择提供了一个全面的工具包，以标记和可视化各种分子的利益与精致的特异性和高灵敏度。虽然荧光显微镜目前是细胞成像的首选方法，但它在研究大量小生物分子时面临着根本性的限制。这是因为相对庞大的普通荧光标记可能会引入相当大的扰动，甚至完全改变重要小生物分子的天然功能。因此，尽管它们具有巨大的功能重要性，但这些小生物分子在很大程度上仍然无法通过荧光显微镜检测到。为了应对这一挑战，最近推出了生物正交化学成像平台。受激拉曼散射（SRS）显微术是一种新兴的非线性拉曼显微术，它通过耦合微小的拉曼活性振动探针（如：G.生物正交化学成像技术（Bioorthogonal Chemical Imaging）具有卓越的灵敏度、特异性和生物相容性，可用于活体系统中的小生物分子成像。在这个账户中，我们回顾了最近的技术成就，用于可视化广谱的小生物分子，包括核糖核苷/脱氧核糖核苷，氨基酸，脂肪酸，胆碱，葡萄糖，胆固醇和小分子药物在活的生物系统，从单个细胞到动物组织和模型生物体。重要的是，该平台与活细胞生物学兼容，从而允许对小分子动力学进行实时成像。此外，我们讨论了进一步的化学和光谱策略，生物正交化学成像，一个有价值的技术在“组学”的时代。作为生物学发现的独特工具，该平台已被应用于研究生理和病理状态下的各种代谢过程，包括神经系统的蛋白质合成活性、亨廷顿病模型中的蛋白质聚集、肿瘤异种移植物中的葡萄糖摄取以及药物通过皮肤组织的渗透。我们设想，SRS显微镜与振动探针的耦合将对小的生物分子做荧光团的荧光显微镜对较大的分子种类做的事情。
Innovations in light microscopy have tremendously revolutionized the way researchers study biological systems with subcellular resolution. In particular, fluorescence microscopy with the expanding choices of fluorescent probes has provided a comprehensive toolkit to tag and visualize various molecules of interest with exquisite specificity and high sensitivity. Although fluorescence microscopy is currently the method of choice for cellular imaging, it faces fundamental limitations for studying the vast number of small biomolecules. This is because that common fluorescent labels, which are relatively bulky, could introduce considerable perturbation to or even completely alter the native functions of vital small biomolecules. Hence, despite their immense functional importance, these small biomolecules remain largely undetectable by fluorescence microscopy. To address this challenge, a Bioorthogonal Chemical Imaging platform has recently been introduced. By coupling the stimulated Raman scattering (SRS) microscopy, an emerging nonlinear Raman microscopy technique, with tiny and Raman-active vibrational probes (e. g. alkynes and stable isotopes), Bioorthogonal Chemical Imaging exhibits superb sensitivity, specificity and biocompatibility for imaging small biomolecules in live systems. In this Account, we review recent technical achievements for visualizing a broad spectrum of small biomolecules, including ribonucleosides/deoxyribonucleosides, amino acids, fatty acids, choline, glucose, cholesterol and small-molecule drugs in live biological systems ranging from individual cells to animal tissues and to model organisms. Importantly, this platform is compatible with live-cell biology, thus allowing real-time imaging of small-molecule dynamics. Moreover, we discuss further chemical and spectroscopic strategies for multicolor Bioorthogonal Chemical Imaging, a valuable technique in the era of “omics”. As a unique tool for biological discovery, this platform has been applied to studying various metabolic processes under both physiological and pathological states, including protein synthesis activity of neuronal systems, protein aggregations in Huntington disease models, glucose uptake in tumor xenograft and drug penetration through skin tissues. We envision that the coupling of SRS microscopy with vibrational probes would do for small biomolecules what fluorescence microscopy of fluorophore has done for larger molecular species.
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影响因子：
18.3
作者：
Hang, Howard C.;Wilson, John P.;Charron, Guillaume
通讯作者：
Charron, Guillaume
影响因子：
3.3
作者：
Fu, Dan;Holtom, Gary;Freudiger, Christian;Zhang, Xu;Xie, Xiaoliang Sunney
通讯作者：
Xie, Xiaoliang Sunney
DOI：
10.1039/c3an02281a
发表时间：
2014-05-21
期刊：
The Analyst
影响因子：
--
作者：
Hu F;Wei L;Zheng C;Shen Y;Min W
通讯作者：
Min W
影响因子：
15
作者：
Chen, Zhixing;Paley, Daniel W.;Wei, Lu;Weisman, Andrew L.;Friesner, Richard A.;Nuckolls, Colin;Min, Wei
通讯作者：
Min, Wei
DOI：
10.1073/pnas.0907864106
发表时间：
2009-09-08
影响因子：
11.1
作者：
Jao, Cindy Y.;Roth, Mary;Salic, Adrian
通讯作者：
Salic, Adrian