Spatial distribution of the state of water in frozen mammalian cells

Jinping Dong, Jason Malsam, John C. Bischof, Allison Hubel, Alptekin Aksan

Research output: Contribution to journalArticlepeer-review

40 Scopus citations

Abstract

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 × 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. These observations have a very significant impact for developing successful biopreservation protocols for cells used for therapeutic purposes and for cellular biofluids.

Original languageEnglish (US)
Pages (from-to)2453-2459
Number of pages7
JournalBiophysical journal
Volume99
Issue number8
DOIs
StatePublished - Oct 20 2010

Bibliographical note

Funding Information:
Funding for this research was provided by University of Minnesota Biopreservation Core Resource (BioCoR), an entity funded by the Academic Health Center, College of Science and Engineering, Medical School, Medical Devices Center, and the Institute for Engineering in Medicine at the University of Minnesota. Partial funding for pilot experiments was supplied by the Biotransport Center at the University of Minnesota.

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