New violet-excitable reagents for multicolor flow applications


  • While exploratory work has been done on an improved AmCyan (AmCyan100; [15]), V500 has been developed into products while AmCyan100 is only available through collaborative arrangements. V500 and AmCyan100 are both desirable because of their spectral characteristics, which should make either one suitable for use in combination with other violet-excitable dyes such as Alexa Fluor® 405, BV421, V450, and Pacific Blue.

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We have recently added three new fluorophores–BD Horizon™ V450, BD Horizon V500, and BD Horizon V550 (V450, V500, and V550; BD Biosciences, San Jose, CA) to our existing AmCyan product, forming a group of four violet-excitable dyes from which we have produced functional antibody conjugates. These conjugates, with emission maxima that range from 450 to 535 nm, are compatible with multilaser flow cytometry (FCM) and can be used for polychromatic FCM in three-color or two-color combinations; in fact, V500 fills a spectral opening that has thus far not been exploited by other manufacturers of FCM reagents. We here report that conjugates based on BD Horizon dyes performed well within a useful sensitivity range, established by testing a representative group of violet-excitable FCM reagents currently available, and that V500 has better compatibility with FITC in multicolor applications than does AmCyan. © 2013 International Society for Advancement of Cytometry