Reduced background autofluorescence for cell imaging using nanodiamonds and lanthanide chelates.
Reduced background autofluorescence for cell imaging using nanodiamonds and lanthanide chelates.
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DOI：
10.1038/s41598-018-22702-1
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发表时间：
2018-03-14
影响因子：
4.6
通讯作者：
Packer NH
中科院分区：
文献类型：
--
作者：
Cordina NM;Sayyadi N;Parker LM;Everest-Dass A;Brown LJ;Packer NH
生物成像是跟踪和监测重要生物过程和基本生物分子相互作用的关键技术，然而背景自体荧光与靶向荧光团的干扰对于许多生物成像应用是有问题的。本研究报告了两种新的方法，以减少干扰细胞自发荧光的生物成像。第一种方法使用含有氮空位中心的荧光纳米金刚石（FND）。FND在通常高于大多数细胞自发荧光的近红外波长下发射;并且当适当官能化时，可用于靶向生物分子的无背景成像。第二种方法使用具有长荧光寿命的铕螯合标签。由于细胞自发荧光的短荧光寿命，这些铕螯合标签增强了无背景成像。在这项研究中，我们使用这两种方法来靶向E-选择素，一种跨膜糖蛋白，被炎症激活，以证明在固定的内皮细胞无背景荧光染色。我们的研究结果表明，FND和铕基染色都可以通过减少与细胞自发荧光的竞争来提高荧光生物成像能力。发现涂覆有E-选择素抗体的30 nm纳米金刚石能够最灵敏地检测发炎细胞中的E-选择素，检测到的强度增加40倍。
Bio-imaging is a key technique in tracking and monitoring important biological processes and fundamental biomolecular interactions, however the interference of background autofluorescence with targeted fluorophores is problematic for many bio-imaging applications. This study reports on two novel methods for reducing interference with cellular autofluorescence for bio-imaging. The first method uses fluorescent nanodiamonds (FNDs), containing nitrogen vacancy centers. FNDs emit at near-infrared wavelengths typically higher than most cellular autofluorescence; and when appropriately functionalized, can be used for background-free imaging of targeted biomolecules. The second method uses europium-chelating tags with long fluorescence lifetimes. These europium-chelating tags enhance background-free imaging due to the short fluorescent lifetimes of cellular autofluorescence. In this study, we used both methods to target E-selectin, a transmembrane glycoprotein that is activated by inflammation, to demonstrate background-free fluorescent staining in fixed endothelial cells. Our findings indicate that both FND and Europium based staining can improve fluorescent bio-imaging capabilities by reducing competition with cellular autofluorescence. 30 nm nanodiamonds coated with the E-selectin antibody was found to enable the most sensitive detective of E-selectin in inflamed cells, with a 40-fold increase in intensity detected.
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影响因子：
38.3
作者：
Bradac, C.;Gaebel, T.;Rabeau, J. R.
通讯作者：
Rabeau, J. R.
影响因子：
2.8
作者：
Bradac, Carlo;Say, Jana M.;Brown, Louise J.
通讯作者：
Brown, Louise J.
影响因子：
4.4
作者：
Kluger, MS;Shiao, SL;Pober, JS
通讯作者：
Pober, JS
DOI：
10.1007/978-1-60327-198-1_6
发表时间：
2011-01-01
期刊：
CHARACTERIZATION OF NANOPARTICLES INTENDED FOR DRUG DELIVERY
影响因子：
--
作者：
Clogston, Jeffrey D.;Patri, Anil K.
通讯作者：
Patri, Anil K.
影响因子：
20.3
作者：
Luo, JY;Paranya, G;Bischoff, J
通讯作者：
Bischoff, J