This paper describes several techniques and methods for sludge characterization which are recently developed and used in our ongoing research on biological phosphorus removal (BPR): (i) chemical measurement of the poly-hydroxy-alkanoate (PHA) content of the biomass, (ii) bioassay for glycogen determination in BPR systems, (iii) determination of the fraction of denitrifying phosphorus accumulating organisms (DPAOs) in P-removing sludge, (iv) determination of the aerobic and anoxic ATP/NADH2 ratio (δ value) in BPR systems, and (v) determination of the glycogen accumulating organisms (GAOs) / phosphorus accumulating organisms (PAOs) ratio in activated sludge.