This paper might be helpful. 1. J Struct Biol. 2014 Nov;188(2):102-6. doi: 10.1016/j.jsb.2014.09.011. Epub 2014 Oct 5.
Efficient cryoprotection of macromolecular crystals using vapor diffusion of volatile alcohols. Farley C, Juers DH. Author information: (1)Department of Physics, Whitman College, Walla Walla, WA 99362, United States. (2)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: juer...@whitman.edu<mailto:juer...@whitman.edu>. 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. PMCID: PMC4252874 [Available on 2015-11-01] PMID: 25286441 [PubMed - in process] John J. Tanner Professor of Biochemistry and Chemistry Chair, Biochemistry Department Graduate Admissions Committee University of Missouri-Columbia 125 Chemistry Building Columbia, MO 65211 Phone: 573-884-1280 Fax: 573-882-2754 Email: tanne...@missouri.edu<mailto:tanne...@missouri.edu> http://faculty.missouri.edu/~tannerjj/tannergroup/tanner.html On Jun 12, 2015, at 4:11 PM, "Thomas, Leonard M." <lmtho...@ou.edu<mailto:lmtho...@ou.edu>> wrote: Hi All, We have gotten some very nicely formed crystals out of a couple of different volatile solvents recently. Besides looking for something easier to work in does anybody have any tips on handling crystals from these types of solvents. It is very hard to loop a crystal while it is doing the backstroke in the well with all of its buddies. Thanks in advance. Len Leonard M. Thomas Ph.D. Macromolecular Crystallography Laboratory Manager University of Oklahoma Department of Chemistry and Biochemistry Stephenson Life Sciences Research Center 101 Stephenson Parkway Norman, OK 73019 405-325-1126 lmtho...@ou.edu<mailto:lmtho...@ou.edu> http://barlywine.chem.ou.edu http://structuralbiology.ou.edu
