A series of non-ionic organosilicone surfactants with comb structure (PESO) were prepared through hydrosilylation method. The structure of PESO polymers was characterized via infrared spectroscopy and nuclear magnetic resonance hydrogen spectrum. Then, the effects of hydrophobic group content on surface property of PESO surfactant were studied. The adsorption behavior of PESO surfactant on the air/water interface and the aggregation behavior of PESO surfactant in aqueous solution were discussed. The size and distribution of PESO micelle were also investigated. The results show that it is easier for PESO molecules to associate with each other to form micelles with increasing the content of hydrophobic groups. And the average size of micelles increases as well. When the content of PHMS increases from 25% to 67%, the value of CMC decreases from 0.828 g/L to 0.613 g/L. The surface tension is low to 22.11mN/m. Surface adsorption quantity of PESO is high to 3.31 μmol/m². The average size of each surfactant molecules adsorbed on the air/water boundary size is only 0.502 nm². So PESO surfactants have good surface activity. The aggregation behavior of PESO polymers in aqueous solution was studied by fluorescence spectrometry. The results show that （I₁/I₃ value decreases continuously with increasing concentration of surfactant. （I₁/I₃ tends to be a constant when the concentration of PESO reaches its critical association concentration due to pyrene molecules dissolved in PESO micelle.