Labeling enzymes and extracellular polymeric substances in aerobic granules

Multicolor staining technique provides information on how different constituent components distribute in bioaggregates. This work for the first time adopted immunohistochemical staining and the multicolor fluorescent technique to simultaneously examine the spatial distributions of enzymes, extracellular polymeric substances (EPSs) and cells in aerobic granules. Sample tests with granules cultivated from starch or phenol medium were made to demonstrate the feasibility of the proposed scheme. The cross-talk between channels was revealed in table form. The transient staining test proposed the possible mass transfer limit to the applied antibodies. The proposed staining scheme can appropriately probe the distributions of enzymes, EPS and cells in aerobic granules.