Simultaneous Raman microspectroscopy and fluorescence imaging of bone mineralization in living zebrafish larvae.
Simultaneous Raman microspectroscopy and fluorescence imaging of bone mineralization in living zebrafish larvae.
复制标题
DOI：
10.1016/j.bpj.2014.01.002
复制
发表时间：
2014-02-18
影响因子：
3.4
通讯作者：
Masic, A.
中科院分区：
文献类型：
--
作者：
Bennet, M.;Akiva, A.;Faivre, D.;Malkinson, G.;Yaniv, K.;Abdelilah-Seyfried, S.;Fratzl, P.;Masic, A.
共焦拉曼显微光谱和荧光成像是两种成熟的方法，分别提供对细胞外基质和活细胞和组织的功能性洞察，直至单分子检测。然而，在活组织中，细胞和细胞外基质共存并相互作用。为了获得这种细胞-基质相互作用的信息，我们开发了一种共定位，相关的多光谱组织分析技术，通过在共焦拉曼显微镜上实现高灵敏度，宽视场荧光成像。作为一个原则的证明，我们研究骨形成的早期阶段，在斑马鱼（Danio rerio）的幼虫，因为斑马鱼已成为一个模式生物研究脊椎动物的发展。使用钙荧光标记物对新形成的骨进行染色，并对成熟过程进行成像和体内化学表征。从斑马鱼鳍骨中矿物沉积的早期阶段获得的结果明确显示，除了碳酸化磷灰石mineral. The方法开发的代谢活动，细胞内传感，和贩运以及在体内探索（病理）生理条件下的细胞-组织界面的分子成像中打开了重要的机会，磷酸氢盐含矿物相的存在。
Confocal Raman microspectroscopy and fluorescence imaging are two well-established methods providing functional insight into the extracellular matrix and into living cells and tissues, respectively, down to single molecule detection. In living tissues, however, cells and extracellular matrix coexist and interact. To acquire information on this cell-matrix interaction, we developed a technique for colocalized, correlative multispectral tissue analysis by implementing high-sensitivity, wide-field fluorescence imaging on a confocal Raman microscope. As a proof of principle, we study early stages of bone formation in the zebrafish (Danio rerio) larvae because the zebrafish has emerged as a model organism to study vertebrate development. The newly formed bones were stained using a calcium fluorescent marker and the maturation process was imaged and chemically characterized in vivo. Results obtained from early stages of mineral deposition in the zebrafish fin bone unequivocally show the presence of hydrogen phosphate containing mineral phases in addition to the carbonated apatite mineral. The approach developed here opens significant opportunities in molecular imaging of metabolic activities, intracellular sensing, and trafficking as well as in vivo exploration of cell-tissue interfaces under (patho-)physiological conditions.
登录
查看更多内容
影响因子：
3.4
作者：
Klein, Katharina;Gigler, Alexander M.;Schlegel, Juergen
通讯作者：
Schlegel, Juergen
影响因子：
4.1
作者：
Crane, Nicole J.;Popescu, Victoria;Ignelzi, Michael A., Jr.
通讯作者：
Ignelzi, Michael A., Jr.
DOI：
10.1073/pnas.0914218107
发表时间：
2010-04-06
影响因子：
11.1
作者：
Mahamid, Julia;Aichmayer, Barbara;Addadi, Lia
通讯作者：
Addadi, Lia
影响因子：
3.5
作者：
Schulmerich, Matthew V.;Dooley, Kathryn A.;Goldstein, Steven A.
通讯作者：
Goldstein, Steven A.
影响因子：
1.9
作者：
Engel, Sascha Ronald;Koch, Peter;Leipertz, Alfred
通讯作者：
Leipertz, Alfred