Fluorescent avidin-bound silver particle: a strategy for single target molecule detection on a cell membrane.
Fluorescent avidin-bound silver particle: a strategy for single target molecule detection on a cell membrane.
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DOI：
10.1021/ac801932m
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发表时间：
2009-02-01
影响因子：
7.4
通讯作者：
Lakowicz, Joseph R.
中科院分区：
文献类型：
--
作者：
Zhang, Jian;Fu, Yi;Liang, Dong;Zhao, Richard Y.;Lakowicz, Joseph R.
制备了cy5 -亲和素结合银纳米粒子作为荧光分子试剂，用于细胞成像。与不含金属的亲和素缀合物相比，亲和素-金属配合物具有更强的发射强度、更短的寿命和更好的光稳定性。将亲和素-金属配合物与PM1细胞株表面的生物素位点结合，用扫描共聚焦显微镜记录细胞图像。注意到结合在细胞表面的亲和素-金属配合物可以被识别为与细胞自身荧光不同的孤立发射点。随着细胞表面亲和金属配合物数量的增加，细胞图像上的发射强度增加，但寿命降低。在细胞表面亲和金属配合物的数量与整个细胞图像的发射强度或寿命之间实现了定量回归曲线。基于这条曲线，我们期望开发一种方法，可用于在单细胞水平上使用细胞强度和寿命图像来量化细胞表面上目标分子的数量。
Cy5-avidin conjugate-bound silver nanoparticles were prepared as fluorescence molecular reagent for the cell imaging. Compared with the metal-free avidin conjugate, the avidin-metal complex was observed to display a stronger emission intensity, shorter lifetime, and better photostability. The avidin-metal complexes were conjugated with the biotin-sites on the surfaces of PM1 cell lines and the cell images were recorded using scanning confocal microscopy. It was noticed that the avidin-metal complexes bound on the cell surfaces could be identified as the isolated emission spots distinct from the cellular autofluorescence. The emission intensity over the cell image was increased with an increase of the number of avidin-metal complex on the cell surface but the lifetime was decreased. A quantitative regression curve was achieved between the amount of avidin-metal complex on the cell surface and the emission intensity or lifetime over the entire cell image. Based on this curve, we expect to develop an approach that can be used to quantify the amount of target molecules on the cell surfaces using the cell intensity and lifetime images at the single cell level.
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影响因子：
7.4
作者：
Knemeyer, JP;Herten, DP;Sauer, M
通讯作者：
Sauer, M
影响因子：
3.7
作者：
Mertens, H.;Koenderink, A. F.;Polman, A.
通讯作者：
Polman, A.
影响因子：
15
作者：
Hu, J;Zhang, J;Fox, MA
通讯作者：
Fox, MA
影响因子：
3.3
作者：
Bruzzone, S;Malvaldi, M;Guidotti, C
通讯作者：
Guidotti, C
影响因子：
3.9
作者：
Fu, Yi;Zhang, Jian;Lakowicz, Joseph R.
通讯作者：
Lakowicz, Joseph R.