Fluorecent staining for study of extracellular polymeric substances in membrane biofouling layers

Membrane biofouling by microbial products adversely impacts the feasibility of adopting membrane bioreactors (MBRs) for treating wastewater. The fouling layer structure determines the pressure drop across the fouling layer. Three-dimensional distributions of nucleic acids, proteins, alpha-D-GLUCOPYRANOSE POLYSACCHARIDES, AND B-D-glucopyranose polysaccharides in the fouling layer formed on a mixed cellulose ester membrane were generated utilizing a quadruple staining protocol combined with confocal laser scanning microscopy (CLSM). For the first time, this study constructed a three-dimensional volumetric grids model representing the fouling layer structure on the basis of a series of CLSM images. Quantitative structural information about the fouling layer was extracted from the CLSM images.