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In 35 years of negative staining my go to stain is sodium silicotungstate. It provides good contrast has a fine grain, and therefore reveals details not obvious with uranyl salts and is great for really small molecules, it has none of the problems of radioactivity
for handling and disposal. I use it with all the conventional buffers including PBS, but it's of no use when detergent is present. </div>
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UK suppliers sell the acid form (not sure about anywhere else), and to achieve the best affects you need to neutralise your solution. I prefer to prepare a batch of the salt from the acid solution and precipitate it out to keep the solid for later use ... it
keeps for years that way. I then prepare a 1% solution of the salt form, which also keeps for many months. </div>
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If anyone would like the recipe for the conversion to the salt form, I'd be happy to supply it.</div>
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Lesley Calder</div>
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<span style="font-size: 10pt;">The Francis Crick Institute</span><br>
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<div id="divRplyFwdMsg" dir="ltr"><font face="Calibri, sans-serif" style="font-size:11pt" color="#000000"><b>From:</b> 3dem <3dem-bounces@ncmir.ucsd.edu> on behalf of Riedel, Dietmar <driedel@gwdg.de><br>
<b>Sent:</b> 20 October 2021 14:32<br>
<b>To:</b> Sharon G Wolf <Sharon.Wolf@weizmann.ac.il><br>
<b>Cc:</b> 3dem@ncmir.ucsd.edu <3dem@ncmir.ucsd.edu><br>
<b>Subject:</b> Re: [3dem] [EXTERNAL] Uranyl acetate alternative</font>
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<span style="font-size:10.0pt; color:red"><b>External Sender:</span></b> <span style="font-size:10.0pt; color:black">
Use caution.</span></div>
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<div dir="ltr">I use NanoVan or NanoW. NanoVan is great for negativ stain if phosphates omit the use of Uran stains. But it is a vanadium stain and therefore much weaker than Uran salt. I keep it in the fridge for years. </div>
<div dir="ltr">Dietmar <br>
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<div dir="ltr">Mobil gesendet</div>
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<blockquote type="cite">Am 20.10.2021 um 15:07 schrieb Sharon G Wolf <Sharon.Wolf@weizmann.ac.il>:<br>
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<p class="x_MsoNormal"><span style="font-size:11.0pt; font-family:"Calibri",sans-serif; color:#1F497D">I have used sodium silicotungstate on a variety of samples. Also phosphotungstic acid is a classic.
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<p class="x_MsoNormal"><span style="font-size:11.0pt; font-family:"Calibri",sans-serif; color:#1F497D">Also, like Zuben, I have used ammonium molybdate and methylamine tungstate. Both need to be made fresh. They can’t sit on the shelf for a long time like
UAc can.<br>
None of them has as much contrast as UAc, except for the sodium silicotungstate, which can be outstanding. But it can cause some complexes to fall apart.</span></p>
<p class="x_MsoNormal"><span style="font-size:11.0pt; font-family:"Calibri",sans-serif; color:#1F497D">Sharon</span></p>
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<p class="x_MsoNormal"><b><span style="font-size:11.0pt; font-family:"Calibri",sans-serif">From:</span></b><span style="font-size:11.0pt; font-family:"Calibri",sans-serif"> 3dem <3dem-bounces@ncmir.ucsd.edu>
<b>On Behalf Of </b>Brown, Zuben<br>
<b>Sent:</b> Wednesday, 20 October, 2021 15:41<br>
<b>To:</b> Denis Chrétien <denis.chretien@univ-rennes1.fr>; 3dem@ncmir.ucsd.edu<br>
<b>Subject:</b> Re: [3dem] [EXTERNAL] Uranyl acetate alternative</span></p>
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<p class="x_MsoNormal"><span style="font-family:"Calibri",sans-serif; color:black">Hi Denis,</span></p>
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<p class="x_MsoNormal"><span style="font-family:"Calibri",sans-serif; color:black">I've tried ammonium molybdate and methylamine tungstate with acceptable results on different membrane proteins.</span></p>
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<p class="x_MsoNormal"><span style="font-family:"Calibri",sans-serif; color:black">Obviously, the contrast is reduced compared to uranyl acetate but the management of the waste materials is greatly simplified.</span></p>
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<p class="x_MsoNormal"><span style="font-family:"Calibri",sans-serif; color:black">best wishes,</span></p>
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<p class="x_MsoNormal"><span style="font-family:"Calibri",sans-serif; color:black">Zuben</span></p>
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<p class="x_MsoNormal"><b><span style="font-size:11.0pt; font-family:"Calibri",sans-serif; color:black">From:</span></b><span style="font-size:11.0pt; font-family:"Calibri",sans-serif; color:black"> 3dem <<a href="mailto:3dem-bounces@ncmir.ucsd.edu">3dem-bounces@ncmir.ucsd.edu</a>>
on behalf of Denis Chrétien <<a href="mailto:denis.chretien@univ-rennes1.fr">denis.chretien@univ-rennes1.fr</a>><br>
<b>Sent:</b> Wednesday, October 20, 2021 3:35 AM<br>
<b>To:</b> <a href="mailto:3dem@ncmir.ucsd.edu">3dem@ncmir.ucsd.edu</a> <<a href="mailto:3dem@ncmir.ucsd.edu">3dem@ncmir.ucsd.edu</a>><br>
<b>Subject:</b> [EXTERNAL] [3dem] Uranyl acetate alternative</span> </p>
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<p class="x_MsoNormal">Dear all, </p>
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<p class="x_MsoNormal">It becomes difficult to posses Uranyl Acetate salts. Hence we would like to have your feedback concerning alternatives for sections and negative staining. Along this line, we would appreciate your experience concerning this salt :</p>
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<p class="x_MsoNormal"><a href="https://urldefense.proofpoint.com/v2/url?u=https-3A__eur03.safelinks.protection.outlook.com_-3Furl-3Dhttps-253A-252F-252Furldefense.proofpoint.com-252Fv2-252Furl-253Fu-253Dhttps-2D3A-5F-5Fwww.tedpella.com-5FSDS-2D5Fhtml-5F19485-2D5Fsds.pdf-2526d-253DDwMFaQ-2526c-253D-2D35OiAkTchMrZOngvJPOeA-2526r-253DL7-2DzyQ-2D04fFCMRqzLIOnx7H0exGZHwIQe-5FwMPuY600I-2526m-253DhRDyTdJW4hH05tkZJrE0XkfrezWAxeD53hmQWoZC2pk-2526s-253DSzdIxK-5F0wo-2D0i8bTfRNyPQnZX2l3AyLDYJwGNuZsMTs-2526e-253D-26data-3D04-257C01-257C-257C12340787eb834303351c08d993ce1012-257C4eed7807ebad415aa7a99170947f4eae-257C0-257C1-257C637703335522542045-257CUnknown-257CTWFpbGZsb3d8eyJWIjoiMC4wLjAwMDAiLCJQIjoiV2luMzIiLCJBTiI6Ik1haWwiLCJXVCI6Mn0-253D-257C3000-26sdata-3DDKY3YP02zGwZ2rmB7iTw5Ak-252Frg-252BfXZg-252FwaCWZx4LZ40-253D-26reserved-3D0&d=DwMGaQ&c=-35OiAkTchMrZOngvJPOeA&r=L7-zyQ-04fFCMRqzLIOnx7H0exGZHwIQe_wMPuY600I&m=J6SGoWLqZ81NUd74AhpGSKJazdPjFRhsG3xONcgDMBs&s=J9bsJvBcqZ_-NCCpZmjpO7VLh_ZkCV6FZKz7L28CcjI&e=" originalsrc="https://urldefense.proofpoint.com/v2/url?u=https-3A__www.tedpella.com_SDS-5Fhtml_19485-5Fsds.pdf&d=DwMFaQ&c=-35OiAkTchMrZOngvJPOeA&r=L7-zyQ-04fFCMRqzLIOnx7H0exGZHwIQe_wMPuY600I&m=hRDyTdJW4hH05tkZJrE0XkfrezWAxeD53hmQWoZC2pk&s=SzdIxK_0wo-0i8bTfRNyPQnZX2l3AyLDYJwGNuZsMTs&e=" shash="r5JR8KJsy/FtjexYhTCk5nTRZbl2Xq6UA4Eh/qsp5Bk7i391vYQkfg5zE+E6ITkX1SfsbecNkD7qXxej9l8XX1RQPyAk2XR3EhBm1zlaXD4k7ioC5rNqJ6xw/8J6Sj9lOqX7QpQ59bSVYzVDs4latMtvZmQPS0qtwjPf/q3bBJQ=">https://www.tedpella.com/SDS_html/19485_sds.pdf</a></p>
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<p class="x_MsoNormal">Other suggestions are welcomed!</p>
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<p class="x_MsoNormal">Best,</p>
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<p class="x_MsoNormal">Denis</p>
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