dorsal/arxiv
View SchemaEffects of cryoprotectant concentration and cooling rate on vitrification of aqueous solutions
| Authors | Viatcheslav Berejnov, Naji S. Husseini, Osama A. Alsaied, Robert E. Thorne |
|---|---|
| Categories | |
| ArXiv ID | physics/0604109 |
| URL | https://arxiv.org/abs/physics/0604109 |
| DOI | 10.1107/S0021889806004717 |
| Journal | Journal of Applied Crystallography (2006) 39, 244 |
Abstract
Vitrification of aqueous cryoprotectant mixtures is essential in cryopreservation of proteins and other biological samples. We report systematic measurements of critical cryoprotective agent (CPA) concentrations required for vitrification during plunge cooling from T=295 K to T=77 K in liquid nitrogen. Measurements on fourteen common CPAs including alcohols (glycerol, methanol, isopropanol), sugars (sucrose, xylitol, dextrose, trehalose), PEGs (ethylene glycol, PEG 200, PEG 2 000, PEG 20 000), glycols (DMSO, MPD), and salt (NaCl) were performed for volumes ranging over four orders of magnitude from ~nL to 20 mkL, and covering the range of interest in protein crystallography. X-ray diffraction measurements on aqueous glycerol mixtures confirm that the polycrystalline-to-vitreous transition occurs within a span of less than 2% w/v in CPA concentration, and that the form of polycrystalline ice (hexagonal or cubic) depends on CPA concentration and cooling rate. For most of the studied cryoprotectants, the critical concentration decreases strongly with volume in the range from ~5 mkL to ~0.1 mkL, typically by a factor of two. By combining measurements of the critical concentration versus volume with cooling time versus volume, we obtain the function of greatest intrinsic physical interest: the critical CPA concentration versus cooling rate during flash cooling. These results provide a basis for more rational design of cryoprotective protocols, and should yield insight into the physics of glass formation in aqueous mixtures.
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"abstract": "Vitrification of aqueous cryoprotectant mixtures is essential in\ncryopreservation of proteins and other biological samples. We report systematic\nmeasurements of critical cryoprotective agent (CPA) concentrations required for\nvitrification during plunge cooling from T=295 K to T=77 K in liquid nitrogen.\nMeasurements on fourteen common CPAs including alcohols (glycerol, methanol,\nisopropanol), sugars (sucrose, xylitol, dextrose, trehalose), PEGs (ethylene\nglycol, PEG 200, PEG 2 000, PEG 20 000), glycols (DMSO, MPD), and salt (NaCl)\nwere performed for volumes ranging over four orders of magnitude from ~nL to 20\nmkL, and covering the range of interest in protein crystallography. X-ray\ndiffraction measurements on aqueous glycerol mixtures confirm that the\npolycrystalline-to-vitreous transition occurs within a span of less than 2% w/v\nin CPA concentration, and that the form of polycrystalline ice (hexagonal or\ncubic) depends on CPA concentration and cooling rate. For most of the studied\ncryoprotectants, the critical concentration decreases strongly with volume in\nthe range from ~5 mkL to ~0.1 mkL, typically by a factor of two. By combining\nmeasurements of the critical concentration versus volume with cooling time\nversus volume, we obtain the function of greatest intrinsic physical interest:\nthe critical CPA concentration versus cooling rate during flash cooling. These\nresults provide a basis for more rational design of cryoprotective protocols,\nand should yield insight into the physics of glass formation in aqueous\nmixtures.",
"arxiv_id": "physics/0604109",
"authors": [
"Viatcheslav Berejnov",
"Naji S. Husseini",
"Osama A. Alsaied",
"Robert E. Thorne"
],
"categories": [
"physics.bio-ph",
"physics.chem-ph"
],
"doi": "10.1107/S0021889806004717",
"journal_ref": "Journal of Applied Crystallography (2006) 39, 244",
"title": "Effects of cryoprotectant concentration and cooling rate on vitrification of aqueous solutions",
"url": "https://arxiv.org/abs/physics/0604109"
},
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