Malicka Joanna, Gryczynski Ignacy, Lakowicz Joseph R
Center for Fluorescence Spectroscopy, University of Maryland School of Medicine, Department of Biochemistry and Molecular Biology, 725 West Lombard Street, Baltimore, MD 21201, USA.
Biopolymers. 2004 Jun 15;74(3):263-71. doi: 10.1002/bip.20071.
We examined the fluorescent spectral properties of fluorescein-labeled DNA oligomers when directly bound to metallic silver particles via a terminal sulfhydryl group. We found a 12-fold increase in fluorescence intensity and 25-fold decrease in lifetime for a fluorescein residue positioned 23 nucleotides from the silver surface compared to labeled oligomers in free solution. Similar results were found for a 23-mer labeled with five fluorescein residues. The absence of long lifetime components in the intensity decays suggests that all labeled oligomers are bound to silver and affected similarly by the metallic surfaces. These results provide the basic knowledge needed to begin use of metal-enhanced fluorescence for the detection of target sequences in simple formats potentially without a washing separation step. The use of metal-enhanced fluorescence provides a generic approach to obtaining a hybridization-dependent increase in fluorescence with most, if not all, commonly used fluorophores.
我们研究了通过末端巯基直接与金属银颗粒结合的荧光素标记的DNA寡聚物的荧光光谱特性。我们发现,与游离溶液中的标记寡聚物相比,距离银表面23个核苷酸处的荧光素残基的荧光强度增加了12倍,寿命缩短了25倍。对于用五个荧光素残基标记的23聚体也发现了类似的结果。强度衰减中不存在长寿命成分表明所有标记的寡聚物都与银结合,并受到金属表面的类似影响。这些结果提供了开始使用金属增强荧光以简单形式检测靶序列所需的基础知识,可能无需洗涤分离步骤。金属增强荧光的使用提供了一种通用方法,可通过大多数(如果不是全部)常用荧光团获得与杂交相关的荧光增加。