title: Efficient cryoprotection of macromolecular crystals using vapor diffusion of volatile alcohols.
abstract: Macromolecular X-ray crystallography, usually done at cryogenic temperature to limit radiation damage, often requires liquid cryoprotective soaking that can be labor intensive and damaging to crystals. Here we describe a method for cryoprotection that uses vapor diffusion of volatile cryoprotective agents into loop-mounted crystals. The crystal is mounted into a vial containing a small volume of an alcohol-based cryosolution. After a short incubation with the looped crystal sitting in the cryosolution vapor, the crystal is transferred directly from the vial into the cooling medium. Effective for several different protein crystals, the approach obviates the need for liquid soaking and opens up a heretofore underutilized class of cryoprotective agents for macromolecular crystallography.
authors.affiliation: Department of Physics, Whitman College, Walla Walla, WA 99362, United States.
authors.forename: Christopher
authors.initials: C
authors.lastname: Farley
authors.affiliation: Department of Physics, Whitman College, Walla Walla, WA 99362, United States; Program in Biochemistry, Biophysics and Molecular Biology, Whitman College, Walla Walla, WA 99362, United States. Electronic address: juersdh@whitman.edu.
authors.forename: Douglas H
authors.initials: DH
authors.lastname: Juers
mesh_terms: D000438:Alcohols / Q000737:chemistry*; D003451:Cryoprotective Agents / Q000737:chemistry*; D003460:Crystallization / Q000379:methods; D003461:Crystallography / Q000379:methods; D018360:Crystallography, X-Ray / Q000379:methods; D005615:Freezing; D046911:Macromolecular Substances / Q000737:chemistry*
chemical_list: D000438:Alcohols; D003451:Cryoprotective Agents; D046911:Macromolecular Substances
keywords: Cryoprotection; Vapor diffusion; X-ray crystallography