We want to create two populations of alginate spheres, both containing bacterial cells of different origin. We would like to distinguish these populations by previously coloring the alginate with two different dyes. Does anybody have good suggestions on non-diffusive dyes which could be incorporated into alginate which has not formed a gel yet? Preferably these dyes would be non-toxic and give no significant fluorescence in the GFP emission window (~509 nm) upon excitation with 488 nm.
I think you could covalently crosslink a dye containing an amine group (maybe fluoresceinamin) with the carboxylic function of manuronic/guluronic acids of alginate by carbodiimide method.
Great idea Olivier. Unfortunately, Fluorsceinamin has its emission peak at 513 nm which is too close for our GFP application. Would you have other suggestions? Thanks again.
Best
Lars
Haven't done those stainings yet, still gathering experimental protocols. Though may I suggest 2 ref for alginate-like exopolymer staining w/ RUTHENIUM RED, don't know if it works for non gelly alginate, but worth to have a look at:
*Lin et al. 2010, Water Res, 44: 3355-3364.
*Waller et al. 2004, J Microbiol Methods, 58(1): 23-30.
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