Since the discovery of dimethlysulfoxide (DMSO) as a cryoprotective agent in the 1970’s, there have been many efforts to replace this molecule for the preservation of cells. Nature uses combinations of molecules called osmolytes to preserve biological systems against environmental stresses. We hypothesized that combinations of molecules could be used to preserve cells without DMSO. Testing this hypothesis requires the development of a technology platform. One element of the technology platform was low temperature Raman spectroscopy. This label-free technology enables us to understand where protective molecules are and how they interact with water or the cell to protect it. The other technology is an optimization strategy to reduce the experimentation required to optimize a preservation protocol. An algorithm that enables efficient optimization of composition, freezing rate and enables preservation of heterogeneous cell populations will be described. This platform has been used to develop DMSO-free methods of preserving a variety of cell types.