Sample size: an important parameter in flash-cooling macromolecular crystallization solutions

The effect of sample size on flash‐cooling was assessed using an assortment of Hampton Screen I solutions (Hampton Research). These were cryogenically cooled to ascertain the minimum glycerol concentration needed for vitrification of solutions. Solutions were `flash‐cooled' in 1.0, 0.5 and 0.1 ...

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Published in:Journal of applied crystallography 2005-06, Vol.38 (3), p.412-419
Main Authors: Chinte, U., Shah, B., DeWitt, K., Kirschbaum, K., Pinkerton, A. A., Schall, C.
Format: Article
Language:English
Subjects:
Analytical, structural and metabolic biochemistry
Biological and medical sciences
cryocrystallography
flash-cooling
Fundamental and applied biological sciences. Psychology
In solution. Condensed state. Thin layers
Miscellaneous
Molecular biophysics
Physico-chemical properties of biomolecules
Properties of solutions
aggregation and crystallization of macromolecules
Proteins
vitrification
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cited_by cdi_FETCH-LOGICAL-c4552-69e4a5ac5a1084bddc63ca4811e1bd5435d7a07dd68f49295fba3fc1852c2a0c3
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container_issue 3
container_start_page 412
container_title Journal of applied crystallography
container_volume 38
creator Chinte, U.
Shah, B.
DeWitt, K.
Kirschbaum, K.
Pinkerton, A. A.
Schall, C.
description The effect of sample size on flash‐cooling was assessed using an assortment of Hampton Screen I solutions (Hampton Research). These were cryogenically cooled to ascertain the minimum glycerol concentration needed for vitrification of solutions. Solutions were `flash‐cooled' in 1.0, 0.5 and 0.1 mm commercial cryoloops, using a 100 K nitrogen stream generated by an Oxford 600 Cryostream. The loops were visually examined and then X‐ray diffraction data were collected and evaluated for ice formation. The glycerol concentration needed for vitrification was found to decrease significantly as sample size decreased; detailed key data are presented in tabular form. Glass transition temperatures, Tg, were measured for solutions leading to successful vitrification in the 1 mm cryoloop. The trends in the required concentration of glycerol and resultant Tg can be understood through a simplified heat‐transfer analysis. This analysis points to a critical cooling time period for successful vitrification.
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subjects Analytical, structural and metabolic biochemistry
Biological and medical sciences
cryocrystallography
flash-cooling
Fundamental and applied biological sciences. Psychology
In solution. Condensed state. Thin layers
Miscellaneous
Molecular biophysics
Physico-chemical properties of biomolecules
Properties of solutions
aggregation and crystallization of macromolecules
Proteins
vitrification
title Sample size: an important parameter in flash-cooling macromolecular crystallization solutions
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