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Comparison between photostability of Alexa Fluor 448 and Alexa Fluor 647 with conventional dyes FITC

Flow cytometry involves conjugation of an antibody to a fluorochrome and analysis of its expression. The intensity and quality of fluorescence depends on a large number of factors namely antigen density, fluorolabelling techniques, instrument properties, and also on the type of fluorochrome. With the new age of synthetic dyes, better and brighter fluorochromes are constantly added to the existing palette so making the photostaining much simpler. Recent studies have shown that synthetic dyes are brighter and more photostable than the conventional dyes1-6. Mahmoudian Jet al used mouse anti-human nestin monoclonal antibody conjugated to Alexa Fluor 568and Fluorescein isothiocyanate (FITC) dyes andevaluated their photobleaching and photostabilityprofiles by immunohistochemistry and demonstrated that Alexa Fluor dye had brighter fluorescence and more photo-stability than its counterpart FITC7.We intended to explore the photo stability as well as the intensity and resolution of expression of antigens using the conventional dyes and compare these parameters with those of newer synthetic dyes for their prospective use in clinical flow cytometric service.


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