Spatial Distribution of the State of Water in Frozen Mammalian Cells

被引:46
作者
Dong, Jinping [2 ]
Malsam, Jason [1 ]
Bischof, John C. [3 ]
Hubel, Allison [1 ,3 ]
Aksan, Alptekin [1 ,3 ]
机构
[1] Univ Minnesota, Biopreservat Core Resource BioCoR, Minneapolis, MN 55455 USA
[2] Univ Minnesota, Coll Sci & Engn, Characterizat Facil, Minneapolis, MN USA
[3] Univ Minnesota, Dept Mech Engn, Coll Sci & Engn, Minneapolis, MN 55455 USA
基金
美国国家科学基金会;
关键词
VITRIFICATION; VISUALIZATION;
D O I
10.1016/j.bpj.2010.08.035
中图分类号
Q6 [生物物理学];
学科分类号
071011 ;
摘要
We describe direct determination of the state of intracellular water, measurement of the intercellular concentration of a cryoprotectant agent (dimethylsulfoxide), and the distribution of organic material in frozen mammalian cells. Confocal Raman microspectroscopy was utilized at cryogenic temperatures with single live cells to conduct high spatial resolution measurements (350 x 350 x 700 nm), which yielded two, we believe, novel observations: 1), intracellular ice formation during fast cooling (50 degrees C/min) causes more pronounced intracellular dehydration than slow cooling (1 degrees C/min); and 2), intracellular dimethylsulfoxide concentration is lower (by as much as 50%) during fast cooling, decreasing the propensity for intracellular vitrification. These observations have a very significant impact for developing successful biopreservation protocols for cells used for therapeutic purposes and for cellular biofluids.
引用
收藏
页码:2453 / 2459
页数:7
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