Rieger Sandra, Kulkarni Rajan P, Darcy Dan, Fraser Scott E, Köster Reinhard W
GSF - National Research Center for Environment and Health, Institute of Developmental Genetics, Neuherberg-Munich, Germany.
Dev Dyn. 2005 Nov;234(3):670-81. doi: 10.1002/dvdy.20524.
Recently, inorganic fluorescent contrast agents composed of semiconductor materials have been introduced to biological imaging approaches. These so-called quantum dots provide unique and promising properties unreached by organic fluorophores, but their use as contrast agents within live organisms has been limited, probably due in part to concerns about their in vivo tolerance. Using transparent zebrafish embryos, we challenged quantum dots with a series of intravital imaging problems. We show that quantum dots provide a high fluorescent yield within targeted tissues, possess immense photostability, can be targeted to specific subcellular compartments, remain within targeted cells as lineage tracers, are easily separable from conventional organic fluorescent dyes, and are fixable, allowing them to be used in combination with immunohistochemistry after live recordings. Thus, quantum dots combine the specific advantages of different organic fluorescent contrast agents and promise to become the first fluorophore feasible for long-lasting intravital time-lapse studies. Finally, we show by co-labeling blood vessels of the vasculature and major axon tracts of the nervous system that, for establishing these networks, the same guidance cues might be used in some body parts, whereas in others, both networks appear to develop independently from one another. Thus, the bright fluorescence of quantum dots will help to unravel many open questions in the fields of embryology, cell biology, as well as phenotyping and disease diagnosis.
最近，由半导体材料组成的无机荧光造影剂已被引入生物成像方法中。这些所谓的量子点具有有机荧光团所未达到的独特且有前景的特性，但其在活生物体中作为造影剂的应用一直受到限制，这可能部分归因于对其体内耐受性的担忧。利用透明的斑马鱼胚胎，我们用一系列活体成像问题对量子点进行了挑战。我们表明，量子点在靶向组织中具有高荧光产率，具有极大的光稳定性，可以靶向特定的亚细胞区室，作为谱系追踪剂保留在靶向细胞内，易于与传统有机荧光染料分离，并且可以固定，从而使其能够在实时记录后与免疫组织化学结合使用。因此，量子点结合了不同有机荧光造影剂的特定优势，并有望成为首个适用于长期活体延时研究的荧光团。最后，我们通过对脉管系统的血管和神经系统的主要轴突束进行共标记表明，对于建立这些网络，某些身体部位可能使用相同的引导线索，而在其他部位，这两个网络似乎是彼此独立发育的。因此，量子点的明亮荧光将有助于解开胚胎学、细胞生物学以及表型分析和疾病诊断领域中的许多悬而未决的问题。
