Mapping metabolic changes by noninvasive, multiparametric, high-resolution imaging using endogenous contrast.
Mapping metabolic changes by noninvasive, multiparametric, high-resolution imaging using endogenous contrast.
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DOI：
10.1126/sciadv.aap9302
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发表时间：
2018-03
期刊：
影响因子：
13.6
通讯作者：
Georgakoudi I
中科院分区：
文献类型：
--
作者：
Liu Z;Pouli D;Alonzo CA;Varone A;Karaliota S;Quinn KP;Münger K;Karalis KP;Georgakoudi I
双光子成像提供了在单细胞或组织水平上的非侵入性、无标记、定量的代谢变化分析。监测与代谢相关的亚细胞功能和结构变化对于了解健康组织发育和许多疾病的进展至关重要，包括癌症，糖尿病，心血管和神经退行性疾病。不幸的是，为此目的建立的方法要么是破坏性的，要么需要使用外源性试剂。最近的工作强调了内源性双光子激发荧光（TPEF）作为监测微妙代谢变化的方法的潜力;然而，对检测到的光学信号与潜在代谢途径之间的联系缺乏机械理解。我们提出了一种定量的方法来检测功能和结构代谢生物标志物的非侵入性，依赖于内源性TPEF从两个辅酶，NADH（还原形式的烟酰胺腺嘌呤二核苷酸）和FAD（黄素腺嘌呤二核苷酸）。我们对完整的活细胞和三维组织内的三种光学生物标志物进行多参数分析：细胞氧化还原状态，NADH荧光寿命和线粒体聚类。我们监测受到代谢扰动的细胞和组织中的生物标志物，这些代谢扰动触发不同代谢过程的变化，包括糖酵解和β-内酰胺分解、外源性和内源性线粒体解偶联以及脂肪酸氧化和合成。我们证明，这些光学生物标志物提供了互补的见解的基础生物学机制。因此，当组合使用时，这些生物标志物可以作为敏感的、无标记的鉴定特定代谢途径的变化和表征具有单细胞分辨率的引起的应答的异质性的有价值的工具。
Two-photon imaging provides noninvasive, label-free, quantitative assays of metabolic changes at the single-cell or tissue level. Monitoring subcellular functional and structural changes associated with metabolism is essential for understanding healthy tissue development and the progression of numerous diseases, including cancer, diabetes, and cardiovascular and neurodegenerative disorders. Unfortunately, established methods for this purpose either are destructive or require the use of exogenous agents. Recent work has highlighted the potential of endogenous two-photon excited fluorescence (TPEF) as a method to monitor subtle metabolic changes; however, mechanistic understanding of the connections between the detected optical signal and the underlying metabolic pathways has been lacking. We present a quantitative approach to detecting both functional and structural metabolic biomarkers noninvasively, relying on endogenous TPEF from two coenzymes, NADH (reduced form of nicotinamide adenine dinucleotide) and FAD (flavin adenine dinucleotide). We perform multiparametric analysis of three optical biomarkers within intact, living cells and three-dimensional tissues: cellular redox state, NADH fluorescence lifetime, and mitochondrial clustering. We monitor the biomarkers in cells and tissues subjected to metabolic perturbations that trigger changes in distinct metabolic processes, including glycolysis and glutaminolysis, extrinsic and intrinsic mitochondrial uncoupling, and fatty acid oxidation and synthesis. We demonstrate that these optical biomarkers provide complementary insights into the underlying biological mechanisms. Thus, when used in combination, these biomarkers can serve as a valuable tool for sensitive, label-free identification of changes in specific metabolic pathways and characterization of the heterogeneity of the elicited responses with single-cell resolution.
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影响因子：
7.4
作者：
Blacker, Thomas S.;Duchen, Michael R.
通讯作者：
Duchen, Michael R.
DOI：
10.1152/ajpendo.00117.2011
发表时间：
2012-05-01
影响因子：
5.1
作者：
De Pauw, Aurelia;Demine, Stephane;Arnould, Thierry
通讯作者：
Arnould, Thierry
影响因子：
23.9
作者：
Brooks MD;Burness ML;Wicha MS
通讯作者：
Wicha MS
无标记的体内细胞级检测和凋亡的成像。
影响因子：
2.8
作者：
Bower, Andrew J.;Marjanovic, Marina;Zhao, Youbo;Li, Joanne;Chaney, Eric J.;Boppart, Stephen A.
通讯作者：
Boppart, Stephen A.
影响因子：
4.6
作者：
Alonzo CA;Karaliota S;Pouli D;Liu Z;Karalis KP;Georgakoudi I
通讯作者：
Georgakoudi I