Vorob'ev I A, Rafalovskaia-Orlovskaia E P, Gladkikh A A, Potashnikova D M, Barteneva N S
Tsitologiia. 2011;53(5):392-403.
Quantum dots (QD) nanocrystals consisting of CdSe core with ZnS shell are a novel class of fluorophores with tremendous potential in microscopy and cytometry techniques. The unique optical features of Qdots, namely, high photostability and extinction coefficient, wide absorption and narrow emission spectra, and large Stokes shift make them desirable fluorescent tags for diverse biomedical applications. Applications of this novel technology in microscopy and cytometry produce reliable multicolor specimens due to increased photostability, ability for multiplexing and narrow emission spectra of nanocrystals. QD conjugates are available on the market and could be prepared in the laboratory. This paper describes the application of QD-conjugates for immunophenotyping and FISH assessment of cells and tissues, and the requirements for microscope and flow cytometer reengineering for successful use of QD in multiplex fluorescent format. Despite the considerable progress, important methodological issues still need to be solved in terms of QD nanocrystals' size, heterogeneity, functionalization and stability of their conjugates. We discuss practical approaches and challenges that need to be addressed to make QD immunostaining a standard method in biology.
由CdSe核和ZnS壳组成的量子点（QD）纳米晶体是一类新型荧光团，在显微镜和细胞计数技术方面具有巨大潜力。量子点独特的光学特性，即高光稳定性和消光系数、宽吸收光谱和窄发射光谱以及大斯托克斯位移，使其成为适用于多种生物医学应用的理想荧光标记。由于光稳定性提高、可复用性以及纳米晶体的窄发射光谱，这项新技术在显微镜和细胞计数中的应用产生了可靠的多色样本。量子点偶联物在市场上有售，也可以在实验室制备。本文描述了量子点偶联物在细胞和组织免疫表型分析及荧光原位杂交（FISH）评估中的应用，以及为成功以多荧光形式使用量子点而对显微镜和流式细胞仪进行改造的要求。尽管取得了相当大的进展，但在量子点纳米晶体的尺寸、异质性、功能化及其偶联物的稳定性方面，仍有重要的方法学问题需要解决。我们讨论了为使量子点免疫染色成为生物学中的标准方法而需要应对的实际方法和挑战。
