This material was originally published in the Purdue Cytometry CD-ROM Series

Step 3: Gating strategy To Discriminate Indigenous Microflora And Soil Particles From Sphingomonas Sp. 107.

Bacteria were extracted from soil, permeabiliized and hybridized with fluorescein-labelled specific or non-specific probes. The acquisition regions were defined by FCM analysis of pure fluorescent Sphinomonas sp. 107 cells hybridized with the specific probe (A). First, the scattered region was delimited (region A) and its green fluorescence measured by the FL1 detector (region b). Second, among green fluorescent events which have been detected, red EtBr fluorescent bacteria were discriminated using the FL3 detector (region E). Finally, bacteria stained by both fluorescein labelled oligonucleotide and EtBr were quantified (region H).

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