Hi Thomas, This is usual when high volatile solvent is used in crystallization (membrane or glycoproteins). The crystal may looks very nice with sharp edges. When you open the cover glass, you may see a very thin film formed on the hang-on drops. Once you touch the drop, then crystals move very quickly like flying and start cracking.
Here is what you may try: 1. When you open the cover glass, using a big drop of Paratone oil to cover (wrap) the drop immediately and completely. 2. Then, inject the cryoprotectant into the drop wrapped by paratone oil. 3. When you wash the crystal in cryoprotectant and fish the crystal out, please always keep all of the operation in a large paratone oil drop. 4. Make sure there is always a layer of paratone oil on the loop when you freeze it in LN2. I hope this will be helpful. Regards, Kevin On Tue, Aug 14, 2018 at 12:00 PM Thomas Krey <krey.tho...@mh-hannover.de> wrote: > 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 > > > > ------------------------------ > > To unsubscribe from the CCP4BB list, click the following link: > https://www.jiscmail.ac.uk/cgi-bin/webadmin?SUBED1=CCP4BB&A=1 > -- Kevin Jin Sharing knowledge each other is always very joyful...... Website: http://www.jinkai.org/ ######################################################################## To unsubscribe from the CCP4BB list, click the following link: https://www.jiscmail.ac.uk/cgi-bin/webadmin?SUBED1=CCP4BB&A=1
