Spatial Distribution of the State of Water in Frozen Mammalian Cells
Published on: Tuesday 02, November 2010Doug J, Malsam J, Bischof JC, Hubel A, Aksan A, "Spacial distribution of the state of water in frozen mammalian cells, Biophys J, 99: 2453-2459, 2010.
The direct determination of the state of intracellular water, mearsurement of the intercellular concentration of a cryoprotectant agent (dimethylsulfoxide), and the distribution of organic material in frozen mammalian cells is described. Confocal Raman micrspectroscopy was utilized at cryogenic temperatures with single live cells to conduct high spatial resolution measurements (350_350_700 nm), which yielded two, we believe, novel observations: 1), intracellular ice formation during fast cooling (50_C/min) causes more pronounced intracellular dehydration than slow cooling (1_C/min); and 2), intracellular dimethylsulfoxide concentration is lower (by as much as 50%) during fast cooling, decreasing the propensity for intracellular vitrification.
