File:Cluster of PAOs.jpg

English: Intracellularly stored poly phosphorus (Poly-P) and polyhydroxy-alkanoates (PHA) are the primary targets used to identify phosphorus accumulating organisms (PAOs). The detection of intercellularly stored Poly-P itself is far more reliable than PHA. Polymer staining techniques such as 41,6-diamidino-2-phenylindole (DAPI) are used to identify the presence of PAOs within wastewaters activated sludge. DAPI, a dying molecule, exhibits a maximum fluorescence emission of around 450 nm (blue), when bound to Poly-P, a bathochromic shift in the emission to 525-550 nm occurs, exhibiting a yellowish color. DAPI stains polymeric ions, such as DNA and lipids. DAPI-DNA fluorescence appears blue, whereas both DAPI-Poly-P and DAPI-lipids resonate yellow. To differentiate between the DAPI-poly-P and DAPI-lipids, biomass samples are collected under aerobic conditions where PHA reserves are thought to have been thoroughly consumed. Any yellow dots in this image represent microorganisms that have consumed orthophosphate as a means for binary fission. DAPI-poly-P interaction is extraordinarily precise with the ability to generate an accurate quantum yield from microscopic observation.