荧光标准微球的使用成为定量流式细胞术的标准品。他能够直观的对比不同实验室数据，同临床流式细胞仪上的质量控制一样的好。在这篇文章中，我们描述的一个简单的方法，为生产基于链霉亲和素功能量子点的多色浓缩标准微球。根据他们广阔的吸收光谱和相对较窄的发射光谱，这可以通过斑点大小调节，量子点标准微球可以提供任何荧光相匹配的发射光。早先公布了，我们具有光学特征的商业化链霉亲和素功能量子点。本文中我们描述了，在生物素小球中这些点是如何组装的。应用质量作用定律可以很容易的确定在小球中这些点的密度。这些小球的特性已经通过测试行业标准，即商业化的荧光标准微球。校准微球的应用已经扩展到外皮生长因子配合基表面密度的测定以及点标记的细胞病毒微粒结合物的定量分析。
The development of quantum dot calibration beads and quantitative multicolor bioassays in flow cytometry and microscopy
Yang Wu a,b, Samuel K. Campos c, Gabriel P. Lopez b, Michelle A. Ozbun c,
Larry A. Sklar a, Tione Buranda a,*
a Department of Pathology and Cancer Center, University of New Mexico, Albuquerque, NM 87131, USA
b Department of Chemical and Nuclear Engineering, University of New Mexico, Albuquerque, NM 87131, USA
c Department of Molecular Genetics and Microbiology, University of New Mexico, Albuquerque, NM 87131, USA
Received 22 December 2006
Available online 13 February 2007
Abstract
The use of fluorescence calibration beads has been the hallmark of quantitative flow cytometry. It has enabled the direct comparison of interlaboratory data as well as quality control in clinical flow cytometry. In this article, we describe a simple method for producing color-generalizable calibration beads based on streptavidin functionalized quantum dots. Based on their broad absorption spectra and relatively narrow emission, which is tunable on the basis of dot size, quantum dot calibration beads can be made for any fluorophore that matches their emission color. In an earlier publication, we characterized the spectroscopic properties of commercial streptavidin functionalized dots (Invitrogen). Here we describe the molecular assembly of these dots on biotinylated beads. The law of mass action is used to readily define the site densities of the dots on the beads. The applicability of these beads is tested against the industry standard, namely commercial fluorescein calibration beads. The utility of the calibration beads is also extended to the characterization surface densities of dot-labeled epidermal growth factor ligands as well as quantitative indicators of the binding of dot-labeled virus particles to cells. 2007 Elsevier Inc. All rights reserved.
Keywords: Nanotechnology; Quantum dots; Calibration beads; Flow cytometer; Quantitation; Fluorescence; Multiplex; Bioassays; Virus particles; Microscopy; Spectroscopy; Mass action
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