MiTeGen have instructions on their website for using their system. In practice I usually put a little silicon grease at the base of the capillary as sometimes the capillaries fall off plus you need reasonable steady hand eye coordination to put the capillary over the loop...I have found MiTeGen RT system very useful.
Good luck! -----Original Message----- From: CCP4 bulletin board [mailto:ccp...@jiscmail.ac.uk] On Behalf Of Ezra Peisach Sent: Wednesday, January 27, 2010 7:41 AM To: CCP4BB@JISCMAIL.AC.UK Subject: Re: [ccp4bb] Crystal rescue MiTeGen (www.mitegen.com) sells a RT device - plastic capillary to put over loop... I do not know how well it would work for a long data collection - but people here have used it to evaluate their crystals.... Ezra On 1/27/2010 3:58 AM, Flip Hoedemaeker wrote: > Zhiyi, > > You can use a thin cap over your cryo loop, just put a drop of mother > liquor in the top, place over the loop and make it airtight at the > base. Not sure who sells these things though, I guess you can make it > from a capillary too. Then remove the cryo stream or put it at a temp > above freezing, say 253K. > > Flip > > Zhiyi Wei wrote: >> Thanks for so many quick responses! >> >> Actually, I have test several different cryo-protectants, including >> glycerol, EG, and PEG400. I did not see much differences between these >> cryo conditions. So, I choose glycerol. >> >> I would like to test my crystals in RT. But I don't know how to do >> this. Just mount crystal to the X-ray machine without cryo stream? Or >> I should use capillaries? >> >> Zhiyi >> >> On Wed, Jan 27, 2010 at 5:45 AM, Kelly Daughtry <kddau...@bu.edu> wrote: >>> Tascimate can be used as the cryo as well. I have had experience with >>> crystals in similar condition and moved the crystals to a 20% >>> increased Tascimate solution and they froze well. >>> >>> I agree with Ezra, room temperature mount your crystal before >>> freezing. It is the only way to know the true problem. >>> >>> >>> Kelly >>> ******************************************************* >>> Kelly Daughtry >>> PhD Candidate >>> Department of Physiology and Biophysics >>> Boston University School of Medicine >>> 590 Commonwealth Ave >>> R 390 >>> Boston MA, 02215 >>> (P) 617-358-5548 >>> ******************************************************* >>> >>> >>> >>> On Tue, Jan 26, 2010 at 1:25 PM, Marcus Winter >>> <marcus.win...@oxford-diffraction.com> wrote: >>>> Dear Zhiyi, >>>> >>>> >>>> Ezra is exactly right, of course. The Oxford Diffraction PX Scanner >>>> system can assess the diffraction qualities of (putative) protein >>>> crystals in situ - in the crystallisation plate. So, directly, you >>>> would discover if your 'big and beautiful' crystals actually diffract >>>> well... in their mother liquor under ambient conditions and before the >>>> addition of any cryo-protect. Do you have a friend or neighbour with >>>> a PX Scanner ? If not, please feel most welcome to contact >>>> Oxford Diffraction: we would be pleased to assist if at all possible. >>>> >>>> >>>> Good Luck and Best Wishes, >>>> >>>> Marcus Winter. >>>> >>>> www.oxford-diffraction.com >>>> >>>> >>>> >>>> >>>> -----Original Message----- >>>> From: CCP4 bulletin board [mailto:ccp...@jiscmail.ac.uk] On Behalf Of >>>> Ezra Peisach >>>> Sent: 26 January 2010 16:01 >>>> To: CCP4BB@JISCMAIL.AC.UK >>>> Subject: Re: [ccp4bb] Crystal rescue >>>> >>>> First you need to establish if it is your cryo conditions or the >>>> crystals. Depending where you are - they might have the equipment >>>> to do >>>> >>>> a wet mount - without freezing. Yes the crystal will not last - but >>>> then you know if the problem is in the >>>> crystal. If it is - you need better crystals. If it is the cryo - >>>> you >>>> need to work on that. Tacsimate is mixture of alot of different >>>> compounds - but the smears are too close together to be a small salt >>>> crystal on top... >>>> >>>> Good luck, >>>> >>>> Ezra >>>> >>>> On 1/26/2010 10:42 AM, Zhiyi Wei wrote: >>>>> Dear all, >>>>> >>>>> I got a problem with my crystals. I have two total different proteins >>>>> that both can be crystallized in the condition with PEG3350 and >>>> Tacsimate >>>>> (although the concentrations are different) with different shapes. >>>>> The >>>>> crystals look big and beautiful. However, when I test them in >>>> synchrotron, >>>>> both of these two types of crystals showed poor diffractions. As >>>> showed in >>>>> the attached diffraction image, the diffraction is up to ~4 A but >>>> smear in >>>>> one direction while<8 A in the other direction. The interesting thing >>>> is >>>>> that the diffraction pattern is similar for all crystals (from two >>>> different >>>>> proteins) that I tested without exception although they belong to >>>> different >>>>> space groups. So, I wonder whether these kind of pattern is caused by >>>>> Tacsimate (I don't know what it is) and how to rescue these crystals. >>>> Any >>>>> suggestions or comments? >>>>> >>>>> Thanks a lot! >>>>> >>>>> Best, >>>>> Zhiyi >>>>> >> Notice: This e-mail message, together with any attachments, contains information of Merck & Co., Inc. (One Merck Drive, Whitehouse Station, New Jersey, USA 08889), and/or its affiliates Direct contact information for affiliates is available at http://www.merck.com/contact/contacts.html) that may be confidential, proprietary copyrighted and/or legally privileged. It is intended solely for the use of the individual or entity named on this message. 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