Detection of cell and tissue surface antigens using up-converting phosphors: A new reporter technology
Detection of cell and tissue surface antigens using up-converting phosphors: A new reporter technology
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DOI：
10.1006/abio.1998.2965
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发表时间：
1999-02-01
影响因子：
2.9
通讯作者：
Tanke, HJ
中科院分区：
文献类型：
--
作者：
Zijlmans, HJMAA;Bonnet, J;Tanke, HJ
描述了一种用于组织切片或细胞膜抗原敏感检测的新型发光报告器。它由亚微米大小的荧光粉晶体（0.2-0.4 μ m）组成，表面标有亲和素或抗体，能够与完整细胞或组织切片上的抗原特异性结合。这些荧光粉报告体通过将红外光上转换为可见光，表现出双光子的反斯托克斯发光，并被命名为上转换荧光粉技术（UPT）。它们通常由掺有两种不同镧系元素的三氧硫化物组成，在红外激发下表现出光稳定性，可见光（蓝色，绿色和红色）的强发射。本报告描述了将磷颗粒与NeutrAvidin结合，并随后在由组织切片中的前列腺特异性抗原和人淋巴细胞上的CD4膜抗原组成的模型系统中使用这种结合物。采用外延照度荧光显微镜，利用氙灯提供近红外激发，可视化可见发射。UPT的优点是(1)离散波长的永久、强、抗斯托克斯发射；（ii）由于在红外光激发下没有自身荧光，生物标本的对比度不匹配；（iii）同时检测多种目标分析物；（四）低成本的显微镜改造。这种新方法不仅在诊断病理学方面具有很高的潜在价值，而且在分子生物学、基因组研究、病毒学和微生物学等领域应用时，也可能为蛋白质或核酸的检测提供优势。(C) 1999学术出版社。
A novel luminescent reporter for the sensitive detection of antigens in tissue sections or on cell membranes is described. It consists of submicron-size phosphor crystals (0.2-0.4 mu m), which are surface labeled with avidin or antibodies and capable of binding specifically to antigens on intact cells or in tissue sections. These phosphor reporters exhibit two-photon, anti-Stokes luminescence by up-converting infrared to visible light and are named Up-converting Phosphor Technology (UPT). They typically consist of yttriumoxysulfides doped with two different lanthanides exhibiting photostable, strong emission in the visible (blue, green, and red) upon excitation in the infrared. This report describes the conjugation of phosphor particles to NeutrAvidin with the subsequent use of this conjugate in a model system consisting of prostate-specific antigen in tissue sections and the CD4 membrane antigen on human lymphocytes. An epi-illumination fluorescence microscope was adapted to provide near-IR excitation using a xenon lamp for visualization of the visible emission. Advantages of UPT are (i) permanent, strong, anti-Stokes emission of discrete wavelengths; (ii) unmatched contrast in biological specimens due to the absence of autofluorescence upon excitation with IR light; (iii) simultaneous detection of multiple target analytes; and (iv) low-cost microscope modifications. The new methodology has not only high potential value in diagnostic pathology as described here, but may offer advantages for the detection of proteins or nucleic acids when applied to molecular biology, genomic research, virology, and microbiology. (C) 1999 Academic Press.