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.
Betscheider
Dirk
D
Bioanalytics, Institute of Pharmaceutical and Medicinal Chemistry, Heinrich Heine University, Universitatsstrasse 1, 40225 Düsseldorf, Germany.
Jose
Joachim
J
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