Optical imaging of metabolic dynamics in animals.
Optical imaging of metabolic dynamics in animals.
复制标题
DOI：
10.1038/s41467-018-05401-3
复制
发表时间：
2018-08-06
影响因子：
16.6
通讯作者：
Min W
中科院分区：
文献类型：
--
作者：
Shi L;Zheng C;Shen Y;Chen Z;Silveira ES;Zhang L;Wei M;Liu C;de Sena-Tomas C;Targoff K;Min W
以高空间和时间分辨率直接可视化活体动物的代谢动力学对于理解许多生物过程是必不可少的。在这里，我们介绍了一个结合了氧化氢(D2O)探测和受激拉曼散射(DO-SRS)显微镜来原位成像代谢活动的平台。D2O衍生的大分子在酶作用下会生成碳-氚(C-D)键，这种键可以跟踪组织中的生物合成，并可以用SRS进行原位成像。在C-D键的宽广振动光谱中，我们发现了脂类、蛋白质和DNA特定的拉曼位移，并发展了光谱分解方法来获得具有大分子选择性的C-D信号。DO-SRS显微镜使我们能够探测动物的新生脂肪生成，在没有组织偏见的情况下成像蛋白质的生物合成，并同时可视化脂质和蛋白质代谢并揭示它们的不同动力学。DO-SRS显微镜是一种非侵入性的、普遍适用的、经济实惠的显微镜，可以适用于广泛的生物系统来研究发育、组织动态平衡、衰老和肿瘤异质性。在活体组织中原位成像代谢的非破坏性方法是有限的。在这里，作者结合了氧化氢探测和受激拉曼散射显微镜来成像细胞和体内的脂肪代谢动力学和蛋白质合成，在小鼠、线虫和斑马鱼中。
Direct visualization of metabolic dynamics in living animals with high spatial and temporal resolution is essential to understanding many biological processes. Here we introduce a platform that combines deuterium oxide (D2O) probing with stimulated Raman scattering (DO-SRS) microscopy to image in situ metabolic activities. Enzymatic incorporation of D2O-derived deuterium into macromolecules generates carbon–deuterium (C–D) bonds, which track biosynthesis in tissues and can be imaged by SRS in situ. Within the broad vibrational spectra of C–D bonds, we discover lipid-, protein-, and DNA-specific Raman shifts and develop spectral unmixing methods to obtain C–D signals with macromolecular selectivity. DO-SRS microscopy enables us to probe de novo lipogenesis in animals, image protein biosynthesis without tissue bias, and simultaneously visualize lipid and protein metabolism and reveal their different dynamics. DO-SRS microscopy, being noninvasive, universally applicable, and cost-effective, can be adapted to a broad range of biological systems to study development, tissue homeostasis, aging, and tumor heterogeneity. Non-destructive methods to image metabolism in situ in living tissues are limited. Here the authors combine deuterium oxide probing and stimulated Raman scattering microscopy to image lipid metabolic dynamics and protein synthesis in cells and in vivo in mice, C. elegans, and zebrafish.