Surface occupancy plays a major role in cholesterol's condensing effect.

The condensing power of cholesterol and 5α-cholestane has been examined in liposomal membranes made from 1,2-dipalmitoyl-sn-glycero-3-phosphocholine (DPPC). Quantitative nearest-neighbor recognition (NNR) analysis and fluorescence measurements using phase-sensitive probe Laurdan have demonstrated that 5α-cholestane exhibits a substantially weaker condensing effect. This fact, in and of itself, provides compelling evidence that cholesterol's condensing effect is critically dependent on having its steroid nucleus at the membrane surface.