Fluorescent detection, background reduction

Single molecule detection Counting molecules, rare event detection order of events, probeAarget binding, distribution functions (e.g., number of base pairs for DNA sizing), molecular dynamics Background reduction Fluorescence lifetime allows the identification of a single molecule... 

Affleck RL, Ambrose WP, Demas IN, Goodwin PM, Schecker JA, Wu JM, Keller RA (1996) Reduction of luminescent background in ultrasensitive fluorescence detection by photobleaching. Anal Chem 68(13) 2270-2276. doi 10.1021/ac9512517... 

Reduction of background fluorescence can be achieved by using the fused silica substrate. In this way, the detection limit can be improved [106]. Further reduction in the background could be achieved by optimizing the chip bonding procedure... 

The first automatic immunoanalyzers used fluorescent labels. These provided automation and short turn-around times, but due to a fairly high nonspecific background the sensitivity of assays was not improved over that obtained by RIA. Introduction of time-resolved fluorescence facilitated the development of assays with substantially reduced detection limit [1], here called improved sensitivity. This was based on reduction of the background, which facilitated the use of a large excess of label in sandwich-type immunometric assays and a much extended measuring range [3-5]. Other nomadioactive reporter molecules, e.g., luminescent labels or enzymes combined with luminescent or fluorescent substrates, can provide similar sensitivity of assays [2]. However, time-resolved fluorometry is unique in facilitating development of fully automated in-house assays, and it has therefore become widely used in research laboratories. 

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