Hemoglobin (Hb) and myoglobin (Mb) were encapsulated in transparent silica glasses prepared by the sol-gel method. The preparation of the silica glasses was tailored so that when proteins were entrapped in the pores of the inorganic matrix, they retained their biochemical activity, i.e. they could bind ligands reversibly. Using optical spectroscopy to monitor ligand binding, we studied the binding of O2, CO, and NO with these two heme proteins encapsulated in silica glasses and compared them to heme proteins in aqueous buffer. Both Hb and Mb in the sol-gel glass bound O2, CO, and NO, producing the same spectroscopic properties as those in aqueous buffer. In addition, silica encapsulated Mb was used to evaluate the rate of ligand (O2) transport through the pores of the glass. When varying oxygen concentration and measuring the time required for full conversion of deoxyMb to MbO2 in the silica gel, the time vs. concentration data followed an exponential trend, as expected for diffusion controlled processes.