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<div dir="ltr" data-setdir="false">Dear all, </div><div dir="ltr" data-setdir="false"><br></div><div dir="ltr" data-setdir="false">thank you so much for all your kind help and valuable inputs! </div><div dir="ltr" data-setdir="false"><br></div><div dir="ltr" data-setdir="false"><br></div><div dir="ltr" data-setdir="false">Best regards,<br></div><div dir="ltr" data-setdir="false">Qiaohui</div><div dir="ltr" data-setdir="false"><br></div><div dir="ltr" data-setdir="false"><br></div><div dir="ltr" data-setdir="false"><br></div><div dir="ltr" data-setdir="false"><br></div><div><br></div>
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On Friday, 20 December 2019, 03:31:43 am GMT+8, 3dem-request@ncmir.ucsd.edu <3dem-request@ncmir.ucsd.edu> wrote:
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<div><div dir="ltr">Send 3dem mailing list submissions to<br></div><div dir="ltr"> <a ymailto="mailto:3dem@ncmir.ucsd.edu" href="mailto:3dem@ncmir.ucsd.edu">3dem@ncmir.ucsd.edu</a><br></div><div dir="ltr"><br></div><div dir="ltr">To subscribe or unsubscribe via the World Wide Web, visit<br></div><div dir="ltr"> <a href="https://mail.ncmir.ucsd.edu/mailman/listinfo/3dem" target="_blank">https://mail.ncmir.ucsd.edu/mailman/listinfo/3dem</a><br></div><div dir="ltr">or, via email, send a message with subject or body 'help' to<br></div><div dir="ltr"> <a ymailto="mailto:3dem-request@ncmir.ucsd.edu" href="mailto:3dem-request@ncmir.ucsd.edu">3dem-request@ncmir.ucsd.edu</a><br></div><div dir="ltr"><br></div><div dir="ltr">You can reach the person managing the list at<br></div><div dir="ltr"> <a ymailto="mailto:3dem-owner@ncmir.ucsd.edu" href="mailto:3dem-owner@ncmir.ucsd.edu">3dem-owner@ncmir.ucsd.edu</a><br></div><div dir="ltr"><br></div><div dir="ltr">When replying, please edit your Subject line so it is more specific<br></div><div dir="ltr">than "Re: Contents of 3dem digest..."<br></div><div dir="ltr"><br></div><div dir="ltr"><br></div><div dir="ltr">Today's Topics:<br></div><div dir="ltr"><br></div><div dir="ltr"> 1. Re: Precipitation observed after making freeze substitution<br></div><div dir="ltr"> cocktail containing 5% water (Gibson, Kimberley)<br></div><div dir="ltr"> 2. Re: Precipitation observed after making freeze substitution<br></div><div dir="ltr"> cocktail containing 5% water (Christopher Peddie)<br></div><div dir="ltr"><br></div><div dir="ltr"><br></div><div dir="ltr">----------------------------------------------------------------------<br></div><div dir="ltr"><br></div><div dir="ltr">Message: 1<br></div><div dir="ltr">Date: Thu, 19 Dec 2019 15:08:07 +0000<br></div><div dir="ltr">From: "Gibson, Kimberley" <<a ymailto="mailto:kimberley.gibson@yale.edu" href="mailto:kimberley.gibson@yale.edu">kimberley.gibson@yale.edu</a>><br></div><div dir="ltr">To: Christopher Peddie <<a ymailto="mailto:Christopher.Peddie@crick.ac.uk" href="mailto:Christopher.Peddie@crick.ac.uk">Christopher.Peddie@crick.ac.uk</a>>, Qiaohui Yang<br></div><div dir="ltr"> <<a ymailto="mailto:qiaohui.yang@ymail.com" href="mailto:qiaohui.yang@ymail.com">qiaohui.yang@ymail.com</a>><br></div><div dir="ltr">Cc: "<a ymailto="mailto:3dem@ncmir.ucsd.edu" href="mailto:3dem@ncmir.ucsd.edu">3dem@ncmir.ucsd.edu</a>" <<a ymailto="mailto:3dem@ncmir.ucsd.edu" href="mailto:3dem@ncmir.ucsd.edu">3dem@ncmir.ucsd.edu</a>><br></div><div dir="ltr">Subject: Re: [3dem] Precipitation observed after making freeze<br></div><div dir="ltr"> substitution cocktail containing 5% water<br></div><div dir="ltr">Message-ID:<br></div><div dir="ltr"> <<a ymailto="mailto:BYAPR08MB4886C15DA68DA2CCB921FAF3EB520@BYAPR08MB4886.namprd08.prod.outlook.com" href="mailto:BYAPR08MB4886C15DA68DA2CCB921FAF3EB520@BYAPR08MB4886.namprd08.prod.outlook.com">BYAPR08MB4886C15DA68DA2CCB921FAF3EB520@BYAPR08MB4886.namprd08.prod.outlook.com</a>><br></div><div dir="ltr"> <br></div><div dir="ltr">Content-Type: text/plain; charset="us-ascii"<br></div><div dir="ltr"><br></div><div dir="ltr">It's not possible to make a stock solution of 20% Uranyl Acetate. Even making a 10% UA stock is not possible. At best, the solubility of UA is around 7-8 % in water. It's best to make a 5% stock solution, dissolved in water, for example, and add the quantity of 5%UA to the acetone. The precipitation of UA is likely due to the acetone as UA is not really soluble in acetone.<br></div><div dir="ltr">Best,<br></div><div dir="ltr"><br></div><div dir="ltr">Kimberley Gibson<br></div><div dir="ltr"><br></div><div dir="ltr">Research Associate, Charles Sindelar Lab<br></div><div dir="ltr"><br></div><div dir="ltr">Dept. Molecular Biophysics and Biochemistry<br></div><div dir="ltr">Yale University<br></div><div dir="ltr">________________________________<br></div><div dir="ltr">From: 3dem <<a ymailto="mailto:3dem-bounces@ncmir.ucsd.edu" href="mailto:3dem-bounces@ncmir.ucsd.edu">3dem-bounces@ncmir.ucsd.edu</a>> on behalf of Christopher Peddie <<a ymailto="mailto:Christopher.Peddie@crick.ac.uk" href="mailto:Christopher.Peddie@crick.ac.uk">Christopher.Peddie@crick.ac.uk</a>><br></div><div dir="ltr">Sent: Thursday, December 19, 2019 03:56<br></div><div dir="ltr">To: Qiaohui Yang <<a ymailto="mailto:qiaohui.yang@ymail.com" href="mailto:qiaohui.yang@ymail.com">qiaohui.yang@ymail.com</a>><br></div><div dir="ltr">Cc: <a ymailto="mailto:3dem@ncmir.ucsd.edu" href="mailto:3dem@ncmir.ucsd.edu">3dem@ncmir.ucsd.edu</a> <<a ymailto="mailto:3dem@ncmir.ucsd.edu" href="mailto:3dem@ncmir.ucsd.edu">3dem@ncmir.ucsd.edu</a>><br></div><div dir="ltr">Subject: Re: [3dem] Precipitation observed after making freeze substitution cocktail containing 5% water<br></div><div dir="ltr"><br></div><div dir="ltr"><br></div><div dir="ltr">Hi Qiaohui,<br></div><div dir="ltr"><br></div><div dir="ltr"><br></div><div dir="ltr"><br></div><div dir="ltr">Mix the osmium, water and acetone and cool to -20oC. Then add the uranyl acetate from a stock e.g. 20%. Cooling the mixture in my experience stops the uranyl acetate from precipitating out.<br></div><div dir="ltr"><br></div><div dir="ltr"><br></div><div dir="ltr"><br></div><div dir="ltr">Best,<br></div><div dir="ltr"><br></div><div dir="ltr">Chris<br></div><div dir="ltr"><br></div><div dir="ltr"><br></div><div dir="ltr"><br></div><div dir="ltr"><br></div><div dir="ltr"><br></div><div dir="ltr">From: 3dem [mailto:<a ymailto="mailto:3dem-bounces@ncmir.ucsd.edu" href="mailto:3dem-bounces@ncmir.ucsd.edu">3dem-bounces@ncmir.ucsd.edu</a>] On Behalf Of Qiaohui Yang<br></div><div dir="ltr">Sent: 19 December 2019 08:39<br></div><div dir="ltr">To:<br></div><div dir="ltr">Subject: [3dem] Precipitation observed after making freeze substitution cocktail containing 5% water<br></div><div dir="ltr"><br></div><div dir="ltr"><br></div><div dir="ltr"><br></div><div dir="ltr">Hi everyone!<br></div><div dir="ltr"><br></div><div dir="ltr">We recently tried to make freeze substitution cocktail consisting of 1% osmium tetroxide, 0.1% uranyl acetate and 5% water but observed that the mixture turned slightly cloudy within 1-2 minutes. We tried to see if the precipitation would occur in the absence of osmium tetroxide, and we observed some cloudiness too after mixing uranyl acetate solution in water with pure acetone.<br></div><div dir="ltr"><br></div><div dir="ltr">Just wondering if anyone has encountered similar problem and if so, would appreciate some tips on how to overcome this problem.<br></div><div dir="ltr"><br></div><div dir="ltr">Thank you!<br></div><div dir="ltr"><br></div><div dir="ltr"><br></div><div dir="ltr"><br></div><div dir="ltr">Best regards,<br></div><div dir="ltr"><br></div><div dir="ltr">Qiaohui<br></div><div dir="ltr"><br></div><div dir="ltr"><br></div><div dir="ltr"><br></div><div dir="ltr">The Francis Crick Institute Limited is a registered charity in England and Wales no. 1140062 and a company registered in England and Wales no. 06885462, with its registered office at 1 Midland Road London NW1 1AT<br></div><div dir="ltr">-------------- next part --------------<br></div><div dir="ltr">An HTML attachment was scrubbed...<br></div><div dir="ltr">URL: <<a href="http://mail.ncmir.ucsd.edu/pipermail/3dem/attachments/20191219/ec44c73d/attachment-0001.html" target="_blank">http://mail.ncmir.ucsd.edu/pipermail/3dem/attachments/20191219/ec44c73d/attachment-0001.html</a>><br></div><div dir="ltr"><br></div><div dir="ltr">------------------------------<br></div><div dir="ltr"><br></div><div dir="ltr">Message: 2<br></div><div dir="ltr">Date: Thu, 19 Dec 2019 19:30:52 +0000<br></div><div dir="ltr">From: Christopher Peddie <<a ymailto="mailto:Christopher.Peddie@crick.ac.uk" href="mailto:Christopher.Peddie@crick.ac.uk">Christopher.Peddie@crick.ac.uk</a>><br></div><div dir="ltr">To: "<a ymailto="mailto:3dem@ncmir.ucsd.edu" href="mailto:3dem@ncmir.ucsd.edu">3dem@ncmir.ucsd.edu</a>" <<a ymailto="mailto:3dem@ncmir.ucsd.edu" href="mailto:3dem@ncmir.ucsd.edu">3dem@ncmir.ucsd.edu</a>><br></div><div dir="ltr">Subject: Re: [3dem] Precipitation observed after making freeze<br></div><div dir="ltr"> substitution cocktail containing 5% water<br></div><div dir="ltr">Message-ID:<br></div><div dir="ltr"> <<a ymailto="mailto:VI1PR05MB5726A2F01FAB0FBE282F2FB1D6520@VI1PR05MB5726.eurprd05.prod.outlook.com" href="mailto:VI1PR05MB5726A2F01FAB0FBE282F2FB1D6520@VI1PR05MB5726.eurprd05.prod.outlook.com">VI1PR05MB5726A2F01FAB0FBE282F2FB1D6520@VI1PR05MB5726.eurprd05.prod.outlook.com</a>><br></div><div dir="ltr"> <br></div><div dir="ltr">Content-Type: text/plain; charset="iso-8859-1"<br></div><div dir="ltr"><br></div><div dir="ltr">Sorry, I should have been more specific: 20% in methanol.<br></div><div dir="ltr"><br></div><div dir="ltr">________________________________<br></div><div dir="ltr">From: Gibson, Kimberley <<a ymailto="mailto:kimberley.gibson@yale.edu" href="mailto:kimberley.gibson@yale.edu">kimberley.gibson@yale.edu</a>><br></div><div dir="ltr">Sent: 19 December 2019 15:08<br></div><div dir="ltr">To: Christopher Peddie <<a ymailto="mailto:Christopher.Peddie@crick.ac.uk" href="mailto:Christopher.Peddie@crick.ac.uk">Christopher.Peddie@crick.ac.uk</a>>; Qiaohui Yang <<a ymailto="mailto:qiaohui.yang@ymail.com" href="mailto:qiaohui.yang@ymail.com">qiaohui.yang@ymail.com</a>><br></div><div dir="ltr">Cc: <a ymailto="mailto:3dem@ncmir.ucsd.edu" href="mailto:3dem@ncmir.ucsd.edu">3dem@ncmir.ucsd.edu</a> <<a ymailto="mailto:3dem@ncmir.ucsd.edu" href="mailto:3dem@ncmir.ucsd.edu">3dem@ncmir.ucsd.edu</a>><br></div><div dir="ltr">Subject: Re: [3dem] Precipitation observed after making freeze substitution cocktail containing 5% water<br></div><div dir="ltr"><br></div><div dir="ltr">It's not possible to make a stock solution of 20% Uranyl Acetate. Even making a 10% UA stock is not possible. At best, the solubility of UA is around 7-8 % in water. It's best to make a 5% stock solution, dissolved in water, for example, and add the quantity of 5%UA to the acetone. The precipitation of UA is likely due to the acetone as UA is not really soluble in acetone.<br></div><div dir="ltr">Best,<br></div><div dir="ltr"><br></div><div dir="ltr">Kimberley Gibson<br></div><div dir="ltr"><br></div><div dir="ltr">Research Associate, Charles Sindelar Lab<br></div><div dir="ltr"><br></div><div dir="ltr">Dept. Molecular Biophysics and Biochemistry<br></div><div dir="ltr">Yale University<br></div><div dir="ltr">________________________________<br></div><div dir="ltr">From: 3dem <<a ymailto="mailto:3dem-bounces@ncmir.ucsd.edu" href="mailto:3dem-bounces@ncmir.ucsd.edu">3dem-bounces@ncmir.ucsd.edu</a>> on behalf of Christopher Peddie <<a ymailto="mailto:Christopher.Peddie@crick.ac.uk" href="mailto:Christopher.Peddie@crick.ac.uk">Christopher.Peddie@crick.ac.uk</a>><br></div><div dir="ltr">Sent: Thursday, December 19, 2019 03:56<br></div><div dir="ltr">To: Qiaohui Yang <<a ymailto="mailto:qiaohui.yang@ymail.com" href="mailto:qiaohui.yang@ymail.com">qiaohui.yang@ymail.com</a>><br></div><div dir="ltr">Cc: <a ymailto="mailto:3dem@ncmir.ucsd.edu" href="mailto:3dem@ncmir.ucsd.edu">3dem@ncmir.ucsd.edu</a> <<a ymailto="mailto:3dem@ncmir.ucsd.edu" href="mailto:3dem@ncmir.ucsd.edu">3dem@ncmir.ucsd.edu</a>><br></div><div dir="ltr">Subject: Re: [3dem] Precipitation observed after making freeze substitution cocktail containing 5% water<br></div><div dir="ltr"><br></div><div dir="ltr"><br></div><div dir="ltr">Hi Qiaohui,<br></div><div dir="ltr"><br></div><div dir="ltr"><br></div><div dir="ltr"><br></div><div dir="ltr">Mix the osmium, water and acetone and cool to -20oC. Then add the uranyl acetate from a stock e.g. 20%. Cooling the mixture in my experience stops the uranyl acetate from precipitating out.<br></div><div dir="ltr"><br></div><div dir="ltr"><br></div><div dir="ltr"><br></div><div dir="ltr">Best,<br></div><div dir="ltr"><br></div><div dir="ltr">Chris<br></div><div dir="ltr"><br></div><div dir="ltr"><br></div><div dir="ltr"><br></div><div dir="ltr"><br></div><div dir="ltr"><br></div><div dir="ltr">From: 3dem [mailto:<a ymailto="mailto:3dem-bounces@ncmir.ucsd.edu" href="mailto:3dem-bounces@ncmir.ucsd.edu">3dem-bounces@ncmir.ucsd.edu</a>] On Behalf Of Qiaohui Yang<br></div><div dir="ltr">Sent: 19 December 2019 08:39<br></div><div dir="ltr">To:<br></div><div dir="ltr">Subject: [3dem] Precipitation observed after making freeze substitution cocktail containing 5% water<br></div><div dir="ltr"><br></div><div dir="ltr"><br></div><div dir="ltr"><br></div><div dir="ltr">Hi everyone!<br></div><div dir="ltr"><br></div><div dir="ltr">We recently tried to make freeze substitution cocktail consisting of 1% osmium tetroxide, 0.1% uranyl acetate and 5% water but observed that the mixture turned slightly cloudy within 1-2 minutes. We tried to see if the precipitation would occur in the absence of osmium tetroxide, and we observed some cloudiness too after mixing uranyl acetate solution in water with pure acetone.<br></div><div dir="ltr"><br></div><div dir="ltr">Just wondering if anyone has encountered similar problem and if so, would appreciate some tips on how to overcome this problem.<br></div><div dir="ltr"><br></div><div dir="ltr">Thank you!<br></div><div dir="ltr"><br></div><div dir="ltr"><br></div><div dir="ltr"><br></div><div dir="ltr">Best regards,<br></div><div dir="ltr"><br></div><div dir="ltr">Qiaohui<br></div><div dir="ltr"><br></div><div dir="ltr"><br></div><div dir="ltr"><br></div><div dir="ltr">The Francis Crick Institute Limited is a registered charity in England and Wales no. 1140062 and a company registered in England and Wales no. 06885462, with its registered office at 1 Midland Road London NW1 1AT<br></div><div dir="ltr"><br></div><div dir="ltr">The Francis Crick Institute Limited is a registered charity in England and Wales no. 1140062 and a company registered in England and Wales no. 06885462, with its registered office at 1 Midland Road London NW1 1AT<br></div><div dir="ltr">-------------- next part --------------<br></div><div dir="ltr">An HTML attachment was scrubbed...<br></div><div dir="ltr">URL: <<a href="http://mail.ncmir.ucsd.edu/pipermail/3dem/attachments/20191219/1379e49e/attachment.html" target="_blank">http://mail.ncmir.ucsd.edu/pipermail/3dem/attachments/20191219/1379e49e/attachment.html</a>><br></div><div dir="ltr"><br></div><div dir="ltr">------------------------------<br></div><div dir="ltr"><br></div><div dir="ltr">Subject: Digest Footer<br></div><div dir="ltr"><br></div><div dir="ltr">_______________________________________________<br></div><div dir="ltr">3dem mailing list<br></div><div dir="ltr"><a ymailto="mailto:3dem@ncmir.ucsd.edu" href="mailto:3dem@ncmir.ucsd.edu">3dem@ncmir.ucsd.edu</a><br></div><div dir="ltr"><a href="https://mail.ncmir.ucsd.edu/mailman/listinfo/3dem" target="_blank">https://mail.ncmir.ucsd.edu/mailman/listinfo/3dem</a><br></div><div dir="ltr"><br></div><div dir="ltr"><br></div><div dir="ltr">------------------------------<br></div><div dir="ltr"><br></div><div dir="ltr">End of 3dem Digest, Vol 148, Issue 24<br></div><div dir="ltr">*************************************<br></div></div>
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