Colloidal silver nanoparticles have been synthesized in water-in-oil microemulsion using silver nitrate solubilized in the water core of one microemulsion as source of silver ions, hydrazine hydrate solubilized in the water core of another microemulsion as reducing agent, dodecane as the oil phase, sodium bis(2-ethylhexyl) sulfosuccinate (AOT) as the surfactant. The UV–vis absorption spectra and transmission electron microscopy (TEM) have been used to trace the growth process and elucidate the structure of the silver nanoparticles. UV–vis spectra show that the intermediates formed at early stages of the reaction and then the clusters grow or aggregate to larger nanoparticles. TEM micrographs confirm that the silver nanoparticles are all spherical. The resulting particles have a very narrow size distribution. Meanwhile, the diameter size of the particles is so small that the smallest mean diameter is only 1.6 nm. IR results show that the surfactant molecules are strongly adsorbed on the surface of silver particles through a coordination bond between the silver atom and the sulfonic group of AOT molecules, which endows the particles with a good stability in oil solvents. As dodecane is used as oil solvent to prepare silver nanoparticles, the formed nano-silver sol is almost nontoxic. As a result, the silver nanoparticles need not be separated from the reaction solution and the silver sol may be directly used in antibacterial fields.