Reduction of Luminescent Background in Ultrasensitive Fluorescence Detection by Photobleaching

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• 作者：Rhett L. Affleck ; W. Patrick Ambrose ; James N. Demas ; Peter M. Goodwin ; Jay A. Schecker ; Ming Wu ; and Richard A. Keller
• 刊名：Analytical Chemistry
• 出版年：1996
• 出版时间：July 1, 1996
• 年：1996
• 卷：68
• 期：13
• 页码：2270 - 2276
• 全文大小：174K
• 年卷期：v.68,no.13(July 1, 1996)
• ISSN：1520-6882

文摘

In luminescence-based ultrasensitive analysis, such assingle-molecule detection by flow cytometry, the luminescence background from impurities present in the solventor reagents can ultimately determine the detection limits.A simple, versatile method for reducing luminescencebackground is described. The method is based on photobleaching the reagent stream immediately before itenters the detection flow cell. Dramatic reduction(anorder of magnitude or more) of both low-level continuousbackground and single-molecule fluorescence bursts isdemonstrated. Application and enhancements of thetechnique are discussed.