McGrath Nicole, Barroso Margarida
Albany Medical College, Center for Cardiovascular Sciences, Albany, New York 12208, USA.
J Biomed Opt. 2008 May-Jun;13(3):031210. doi: 10.1117/1.2939417.
Quantum dots (QDs) promise to revolutionize the way fluorescence imaging is used in the cell biology field. The unique fluorescent spectral characteristics, high photostability, low photobleaching, and tight emission spectra of QDs position them above traditional dyes. We will address the ability of water-stabilized QDs to behave as effective fluorescence resonance energy transfer (FRET) donors in cells upon transferrin-receptor-(TFR) mediated endocytosis. Confocal microscopy detects whether donor QD transferrin conjugates transfer energy to acceptor organic fluorophore-transferrin conjugate molecules in endocytic compartments. QDs are shown to be effective FRET donors when internalized into cells via the transferring receptor-mediated endocytic pathway. Upon pairing with the appropriate acceptor dyes, QDs will reduce the laborious data processing that is required to compensate for bleed through contamination between organic dye donor and acceptor pair signals. The QD technology simplifies and expands the use of FRET in the analysis of complex cellular processes that may involve protein organization in intracellular membranes as well as protein-protein interactions.
量子点（QDs）有望彻底改变细胞生物学领域中荧光成像的使用方式。量子点独特的荧光光谱特性、高光稳定性、低光漂白以及狭窄的发射光谱使其优于传统染料。我们将探讨水稳定量子点在转铁蛋白受体（TFR）介导的内吞作用下，在细胞中作为有效荧光共振能量转移（FRET）供体的能力。共聚焦显微镜检测供体量子点 - 转铁蛋白缀合物是否将能量转移到内吞区室中的受体有机荧光团 - 转铁蛋白缀合物分子上。当通过转铁蛋白受体介导的内吞途径内化到细胞中时，量子点被证明是有效的FRET供体。与合适的受体染料配对后，量子点将减少为补偿有机染料供体和受体对信号之间的渗漏污染而所需的繁琐数据处理。量子点技术简化并扩展了FRET在分析复杂细胞过程中的应用，这些过程可能涉及细胞内膜中的蛋白质组织以及蛋白质 - 蛋白质相互作用。
