Fluorescence reference plate for UV illumination using quantum dots.

Abstract

Many image acquisition methods of nucleic acids still depend on UV illumination, especially after the electrophoresis when determining the size of the target DNA. Therefore the quality of the UV illuminator in the gel documentation system, and the comparison of the fluorescence detected are crucial. This paper presents a fluorescence standard reference plate using quantum dots compared to the conventional method where an agarose gel containing ethidium bromide is loaded with standard samples. The fluorescence standard reference plate consists of chambers filled with commercially available quantum dots such as phosphor dots. The chamber is made by thermally attaching nylon and polyester, the former on the inside and the latter on the outside, for increased stability. The images of the proposed reference plate were captured more than 2 months in regular intervals. The intensity analysis of the images shows that the proposed reference plate delivers stable fluorescence over a long term period. The proposed reference plate can be utilized to compare the performance of various UV illuminators, or to set a standard fluorescence point for certain analyses.