I have had some luck with adding ~5-10ul of 60%-70% dilution of the reservoir to the drop. The larger volume of the drop appears to slow down the whizzing of the crystals and allows you to get a few crystals. Though it still occurs. You could also cool the area down or move into the cold room if your crystals survive the transfer as evaporation should be less at 4oC.
You can also spot several 1-2uL of neat reservoir solution around the drop to create a local vapor barrier to prevent evaporation of the drop. It can completely stop the movement of crystals for a few minutes. Dan Daniel A Bonsor PhD. Sundberg Lab Institute of Human Virology University of Maryland, Baltimore 725 W Lombard Street N370 Baltimore Maryland MD 21201 Tel: (410) 706-7457 From: CCP4 bulletin board [mailto:CCP4BB@JISCMAIL.AC.UK] On Behalf Of Thomas Krey Sent: Tuesday, August 14, 2018 2:59 PM To: CCP4BB@JISCMAIL.AC.UK Subject: [ccp4bb] Fishing crystals from volatile solvent as precipitant CAUTION: This message originated from a non UMB, UMSOM, FPI, or UMMS email system. Whether the sender is known or not known, hover over any links before clicking and use caution opening attachments. Dear crystallization experts, We have 3D protein crystals grown from a microseed matrix screening vapor diffusion experiment in either 15% (v/v) Reagent alcohol HEPES Na pH 7.5 0.2 M MgCl2 or in 27% Isopropanol 0.18 M MgCl2 90 mM HEPES Na pH 7.5 10% Glycerol Upon opening the corresponding wells these crystals move quite a bit – presumably due to the volatility of the alcohols. Does anyone have a good suggestion to stabilize the swirling movements? Does anyone have experience, whether these conditions alone can serve as cryo-protectant (i.e., do we really have to fish, move into cryo solution and fish again)? Any suggestion or input would be highly welcome. Thank you very much in advance. Thomas Prof. Dr. Thomas Krey Hannover Medical School Institute of Virology Structural Virology Group Carl-Neuberg-Str. 1 D-30625 Hannover phone: +49 (0) 511 - 532 4308 email: krey.tho...@mh-hannover.de<mailto:krey.tho...@mh-hannover.de> ________________________________ To unsubscribe from the CCP4BB list, click the following link: https://www.jiscmail.ac.uk/cgi-bin/webadmin?SUBED1=CCP4BB&A=1 ######################################################################## To unsubscribe from the CCP4BB list, click the following link: https://www.jiscmail.ac.uk/cgi-bin/webadmin?SUBED1=CCP4BB&A=1
