Multi-Color Single Particle Tracking with Quantum Dots
Multi-Color Single Particle Tracking with Quantum Dots
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DOI：
10.1371/journal.pone.0048521
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发表时间：
2012-11-14
期刊：
影响因子：
3.7
通讯作者：
Lagerholm, B. Christoffer
中科院分区：
文献类型：
--
作者：
Arnspang, Eva C.;Brewer, Jonathan R.;Lagerholm, B. Christoffer
量子点（QD）长期以来一直承诺彻底改变荧光检测，甚至包括需要以单分子灵敏度同时进行多物种检测的应用。尽管早期的承诺，量子点的独特的光学性质尚未得到充分利用，在电子。G.多重单分子灵敏度应用，如单粒子跟踪（SPT）。为了充分优化单分子多重应用与量子点，我们在这项工作中进行了全面的定量调查的荧光强度，荧光强度波动，和水动力学半径的八种类型的市售水溶性量子点。在这项研究中，我们表明，荧光强度的CdSe核心量子点的量子点的发射朝向红色的位移，但混合的CdSe/CdTe核心量子点是不太明亮的最远的红移的CdSe量子点。我们进一步表明，在生物应用中使用蓝移量子点只有一个小的尺寸优势，因为水稳定的表面涂层的额外大小。扩展以前的工作，我们最后还表明，并行的四个彩色双（MC）-SPT与量子点是可能的，在至少25 Hz的图像采集速率。我们通过测量脂质的横向动力学，biotincap-DPPE，在活细胞的细胞质膜使用四种不同颜色的量子点; QD 565，QD 605，QD 655，和QD 705作为标签的技术。
Quantum dots (QDs) have long promised to revolutionize fluorescence detection to include even applications requiring simultaneous multi-species detection at single molecule sensitivity. Despite the early promise, the unique optical properties of QDs have not yet been fully exploited in e. g. multiplex single molecule sensitivity applications such as single particle tracking (SPT). In order to fully optimize single molecule multiplex application with QDs, we have in this work performed a comprehensive quantitative investigation of the fluorescence intensities, fluorescence intensity fluctuations, and hydrodynamic radii of eight types of commercially available water soluble QDs. In this study, we show that the fluorescence intensity of CdSe core QDs increases as the emission of the QDs shifts towards the red but that hybrid CdSe/CdTe core QDs are less bright than the furthest red-shifted CdSe QDs. We further show that there is only a small size advantage in using blue-shifted QDs in biological applications because of the additional size of the water-stabilizing surface coat. Extending previous work, we finally also show that parallel four color multicolor (MC)-SPT with QDs is possible at an image acquisition rate of at least 25 Hz. We demonstrate the technique by measuring the lateral dynamics of a lipid, biotincap-DPPE, in the cellular plasma membrane of live cells using four different colors of QDs; QD565, QD605, QD655, and QD705 as labels.