Design of quantum dot-conjugated lipids for long-term, high-speed tracking experiments on cell surfaces.
Design of quantum dot-conjugated lipids for long-term, high-speed tracking experiments on cell surfaces.
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DOI：
10.1021/ja803325b
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发表时间：
2008-11-12
影响因子：
15
通讯作者：
Naumann, Christoph A.
中科院分区：
文献类型：
--
作者：
Murcia, Michael J.;Minner, Daniel. E.;Mustata, Gina-Mirela;Ritchie, Kenneth;Naumann, Christoph A.
目前的研究报告了量子点（QD）共轭脂质的简便设计及其在细胞表面高速跟踪实验中的应用。 CdSe/ZnS 核/壳量子点，具有两种类型的亲水涂层：2-(2-氨基乙氧基)乙醇（AEE 涂层）和 1,2-二棕榈酰-sn-甘油-3-磷酸胆碱和 60:40 摩尔混合物1,2-二棕榈酰-sn-甘油-3-磷酸乙醇胺-N-[甲氧基（聚乙二醇-2000]（LIPO 涂层），通过 QD 表面上的马来酰亚胺反应基团与巯基脂质缀合。在脂质缀合之前，使用荧光相关光谱法确认了两种类型涂层 QD 在水溶液中的胶体稳定性。基于平面上单脂质跟踪实验的灵敏测定提出了固体支持的磷脂双层，建立了 QD 与脂质的单价缀合的条件，然后将 QD 脂质用作多种细胞类型质膜中的单分子跟踪探针，以 30 fps 的帧速率进行初始跟踪实验，证实 QD 脂质像染料标记的脂质一样在 COS-7、HEK-293、3T3 和 NRK 细胞的质膜中扩散，从而证实了单价。最后，QD-脂质首次应用于高速单分子成像，以高达 1000 fps 的速度跟踪它们在 NRK 成纤维细胞质膜中的横向迁移率。我们的高速跟踪数据与之前使用较大 40 nm Au 标记的跟踪实验非常一致，不仅提高了长时间、连续基于荧光的单分子跟踪的时间分辨率，而且还表明可以采用 10 nm 尺寸的高度光稳定性、光致发光纳米探针。 （AEE 涂层量子点）也很有吸引力，因为与金纳米颗粒不同，它们有助于复杂的多色实验。
The current study reports the facile design of quantum dot (QD)-conjugated lipids and their application to high-speed tracking experiments on cell surfaces. CdSe/ZnS core/shell QDs with two types of hydrophilic coatings, 2-(2-aminoethoxy)ethanol (AEE-coating) and a 60:40 molar mixture of 1,2-dipalmitoyl-sn-glycero-3-phosphocholine and 1,2-dipalmitoyl-sn-glycero-3-phosphoethanolamine-N-[methoxy(polyethyleneglycol-2000] (LIPO-coating), are conjugated to sulfhydryl lipids via maleimide reactive groups on the QD surface. Prior to lipid conjugation, the colloidal stability of both types of coated QDs in aqueous solution is confirmed using fluorescence correlation spectroscopy. A sensitive assay based on single lipid tracking experiments on a planar solid-supported phospholipid bilayer is presented that establishes conditions of monovalent conjugation of QDs to lipids. The QD lipids are then employed as single molecule tracking probes in plasma membranes of several cell types. Initial tracking experiments at a frame rate of 30 fps corroborate that QD-lipids diffuse like dye-labeled lipids in the plasma membrane of COS-7, HEK-293, 3T3, and NRK cells, thus confirming monovalent labeling. Finally, QD-lipids are applied for the first time to high-speed single molecule imaging by tracking their lateral mobility in the plasma membrane of NRK fibroblasts with up to 1000 fps. Our high-speed tracking data, which are in excellent agreement to previous tracking experiments with larger 40nm Au labels, not only push the time resolution in long-time, continuous fluorescence-based single molecule tracking, but also show that highly photostable, photoluminescent nanoprobes of 10nm size can be employed (AEE-coated QDs). These probes are also attractive because, unlike Au nanoparticles, they facilitate complex multicolor experiments.
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影响因子：
46.9
作者：
Lidke, DS;Nagy, P;Jovin, TM
通讯作者：
Jovin, TM
影响因子：
3.4
作者：
Goodwin, JS;Drake, KR;Kenworthy, AK
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2.9
作者：
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作者：
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作者：
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