Abstract: The microstructural properties of gelatin samples derived from the bovine bone as well as skin of fish which inhabit in the cold sea were examined at molecular scale by using the positron annihilation lifetime spectroscopy（PALS）. The longest-lived component of PALS provides information about the free-volume holes in gelatin macromolecules. The results show that both average size and number of free-volume holes in the fish gelatin macromolecules are lower than those in the bovine gelatin macromolecules. Silver halide emulsions were also precipitated by using the balanced double-jet technique，for which two types of media containing fish gelatin and bone gelatin were used in the stage of nucleus formation，respectively. The monodispersibility of silver halide crystallites can be improved and the growth and coalescence of nucleus grains can be restricted when the fish gelatino-peptizer was used as dispersion medium at the nucleus formation. It thus suggested that the function of gelatino-peptizers as protective colloid is correlated with the microstructure character of gelatin macromolecules established by PALS. The gelatino-peptizer derived from the fish skin possesses stronger power governing nucleation and growth of silver halide grains compared with that made from the bovine bone.