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Anal Biochem. 2009 Jan 1;384(1):194-6. doi: 10.1016/j.ab.2008.09.018. Epub 2008 Sep 17.

Nile blue A for staining Escherichia coli in flow cytometer experiments.

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Bioanalytics, Institute of Pharmaceutical and Medicinal Chemistry, Heinrich Heine University, Universitatsstrasse 1, 40225 Düsseldorf, Germany.


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.

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