Nile blue A for staining Escherichia coli in flow cytometer experiments.
Nile blue A is used as a stain for polyhydroxyalkanoic acid-accumulating microorganisms or to detect polyhydroxyalkanoic acids in microorganisms. Here we show that Escherichia coli cells that do not accumulate detectable polyhydroxyalkanoic acids can be stained with Nile blue A and that this staining is sufficient for identifying these cells in fluorescence-activated cell sorting (FACS) experiments. Nile blue A staining did not affect either surface display of peptides or specific labeling of these peptides by a second fluorescence. Staining E. coli for flow cytometry using Nile blue A is an easy-to-handle and low-cost alternative to other fluorescent dyes or the intracellular expression of, for example, green fluorescent protein.
Bioanalytics, Institute of Pharmaceutical and Medicinal Chemistry, Heinrich Heine University, Universitatsstrasse 1, 40225 Düsseldorf, Germany.
Dirk
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Betscheider
Joachim
J
Jose
D002478:Cells, Cultured; D004926:Escherichia coli / Q000378:metabolism*; D005434:Flow Cytometry; D005456:Fluorescent Dyes / Q000737:chemistry*; D049452:Green Fluorescent Proteins / Q000378:metabolism; D010078:Oxazines / Q000737:chemistry*; D013194:Staining and Labeling / Q000379:methods*
D005456:Fluorescent Dyes; D010078:Oxazines; D049452:Green Fluorescent Proteins; C008619:Nile Blue