<html><body><div style="color:#000; background-color:#fff; font-family:garamond, new york, times, serif;font-size:14px"><div id="yui_3_16_0_1_1418998530001_6127" dir="ltr"><span>Hi John and Walter,</span></div><div id="yui_3_16_0_1_1418998530001_6127" dir="ltr"><span><br></span></div><div id="yui_3_16_0_1_1418998530001_6127" dir="ltr"><span id="yui_3_16_0_1_1418998530001_6166">I think that Walter has misnamed the cells. The SedVel50 use the standard width middle rib and have a total of 500 uL space in each sector which uses the 440 and 420 uL to leave a reasonable air space. The SedVel60 cells have a wider middle rib and have smaller total volumes, and I think that it was around 330 and 310 uL for meniscus matching. While I was at Walter's lab we had a few SedVel50's and one SedVel60 for testing, but we still used mostly Walter's homemade meniscus matching cells.</span></div><div id="yui_3_16_0_1_1418998530001_6127" dir="ltr"><span><br></span></div><div id="yui_3_16_0_1_1418998530001_6127" dir="ltr">David</div><br> <div style="font-family: garamond, new york, times, serif; font-size: 14px;" id="yui_3_16_0_1_1418998530001_6107"> <div style="font-family: HelveticaNeue, Helvetica Neue, Helvetica, Arial, Lucida Grande, sans-serif; font-size: 16px;" id="yui_3_16_0_1_1418998530001_6106"> <div dir="ltr" id="yui_3_16_0_1_1418998530001_6126"> <hr size="1" id="yui_3_16_0_1_1418998530001_6139"> <font size="2" face="Arial" id="yui_3_16_0_1_1418998530001_6138"> <b id="yui_3_16_0_1_1418998530001_6439"><span style="font-weight:bold;" id="yui_3_16_0_1_1418998530001_6438">From:</span></b> Walter Stafford <wstafford3@walterstafford.com><br> <b><span style="font-weight: bold;">To:</span></b> John Sumida <jpsumida@U.WASHINGTON.EDU> <br><b><span style="font-weight: bold;">Cc:</span></b> rasmb@rasmb.org <br> <b><span style="font-weight: bold;">Sent:</span></b> Friday, December 19, 2014 2:46 AM<br> <b><span style="font-weight: bold;">Subject:</span></b> Re: [RASMB] Centerpiece question<br> </font> </div> <div class="y_msg_container" id="yui_3_16_0_1_1418998530001_6105"><br><div id="yiv1694068238"><div id="yui_3_16_0_1_1418998530001_6104">Hi John,<div id="yui_3_16_0_1_1418998530001_6103"><span class="yiv1694068238Apple-tab-span" style="white-space:pre;"> </span>The maximum volume depends on whether you are using the SedVel50 or SedVel60 version.You have to fill the sides so that the matched menisci (i.e. the sample and the reference menisci) are both above the capillary (the one that starts on the reference side, runs down the middle of the center rib and into the bottom of the sample side).</div><div id="yui_3_16_0_1_1418998530001_6125">Otherwise, the reference buffer will stop flowing to the sample side when the meniscus gets to the capillary and too little buffer will have been transferred. And the menisci will not be matched.</div><div id="yui_3_16_0_1_1418998530001_6124"><br clear="none"></div><div id="yui_3_16_0_1_1418998530001_6122">You will have to experiment a little to find the sweet spot. I was using the SedVel60 version and - I may not be remembering correctly but I think I used 440 uL on the reference side and 420 uL on the sample side. So that when matched there was 430 uL on each side and my menisci were at 5.90 cm. This gives the longest sedimentation path, if you will. It's wasteful to use a smaller volume: the longer sedimentation path gives you more resolution. Each run is expensive, so you might as well get the most out of it.</div><div><br clear="none"></div><div id="yui_3_16_0_1_1418998530001_6173">Walter<br clear="none"><div class="qtdSeparateBR"><br><br></div><div class="yiv1694068238yqt3692178066" id="yiv1694068238yqt77840"><div><div>On Dec 18, 2014, at 14:23, John Sumida <<a rel="nofollow" shape="rect" ymailto="mailto:jpsumida@U.WASHINGTON.EDU" target="_blank" href="mailto:jpsumida@U.WASHINGTON.EDU">jpsumida@U.WASHINGTON.EDU</a>> wrote:</div><br clear="none" class="yiv1694068238Apple-interchange-newline"><blockquote type="cite"></blockquote></div></div></div></div><div class="yiv1694068238yqt3692178066" id="yiv1694068238yqt34620"><style>#yiv1694068238 #yiv1694068238 --
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#yiv1694068238 </style><div id="yui_3_16_0_1_1418998530001_6179"><div lang="EN-US" id="yui_3_16_0_1_1418998530001_6178"><div class="yiv1694068238WordSection1" id="yui_3_16_0_1_1418998530001_6177"><div class="yiv1694068238MsoNormal"><span style="font-size:12.0pt;">Dear RASMB</span></div><div class="yiv1694068238MsoNormal"><span style="font-size:12.0pt;"> </span></div><div class="yiv1694068238MsoNormal"><span style="font-size:12.0pt;">Thank you for your responses to my question. For the Stafford meniscus matching centerpieces what is the max volume that you can fill these centerpieces to?</span></div><div class="yiv1694068238MsoNormal"><span style="font-size:12.0pt;"> </span></div><div class="yiv1694068238MsoNormal"><span style="font-size:12.0pt;">Best regards,</span></div><div class="yiv1694068238MsoNormal"><span style="font-size:12.0pt;">John Sumida</span></div><div class="yiv1694068238MsoNormal"><span style="font-size:12.0pt;">University of Washington</span></div><div class="yiv1694068238MsoNormal"><span style="font-size:12.0pt;"> </span></div><div id="yui_3_16_0_1_1418998530001_6176"><div style="border:none;border-top:solid #B5C4DF 1.0pt;padding:3.0pt 0in 0in 0in;" id="yui_3_16_0_1_1418998530001_6175"><div class="yiv1694068238MsoNormal" id="yui_3_16_0_1_1418998530001_6174"><b><span style="font-size:10.0pt;">From:</span></b><span style="font-size:10.0pt;"> RASMB [<a rel="nofollow" shape="rect" ymailto="mailto:rasmb-bounces@list.rasmb.org" target="_blank" href="mailto:rasmb-bounces@list.rasmb.org">mailto:rasmb-bounces@list.rasmb.org</a>] <b>On Behalf Of </b>Karl Maluf<br clear="none"><b>Sent:</b> Thursday, December 18, 2014 11:03 AM<br clear="none"><b>To:</b> <a rel="nofollow" shape="rect" ymailto="mailto:rasmb@rasmb.org" target="_blank" href="mailto:rasmb@rasmb.org">rasmb@rasmb.org</a><br clear="none"><b>Subject:</b> Re: [RASMB] Centerpiece question</span></div></div></div><div class="yiv1694068238MsoNormal"> </div><div class="yiv1694068238MsoNormal"><span style="font-size:10.0pt;">The other issue with the meniscus matching centerpeices is to be careful to not overfill the buffer channel higher than the top capillary. This prevents equalization of the pressures between the two channels, and can inhibit the desired meniscus matching.</span><span style="font-size:12.0pt;"></span></div><div><div class="yiv1694068238MsoNormal"><span style="font-size:12.0pt;"> </span></div></div><div><div class="yiv1694068238MsoNormal"><span style="font-size:10.0pt;">Karl</span><span style="font-size:12.0pt;"></span></div></div><div class="yiv1694068238MsoNormal"><span style="font-size:12.0pt;"> </span></div><div align="center" class="yiv1694068238MsoNormal" style="text-align:center;"><span style="font-size:12.0pt;"></span><hr align="center" size="2" width="100%"></div><div class="yiv1694068238MsoNormal" style="margin-bottom:12.0pt;"><b><span style="font-size:10.0pt;">From:</span></b><span style="font-size:10.0pt;"> RASMB [<a rel="nofollow" shape="rect" ymailto="mailto:rasmb-bounces@list.rasmb.org" target="_blank" href="mailto:rasmb-bounces@list.rasmb.org">mailto:rasmb-bounces@list.rasmb.org</a>] <b>On Behalf Of </b>John Philo<br clear="none"><b>Sent:</b> Thursday, December 18, 2014 10:49 AM<br clear="none"><b>To:</b> 'John Sumida'; <a rel="nofollow" shape="rect" ymailto="mailto:rasmb@rasmb.org" target="_blank" href="mailto:rasmb@rasmb.org">rasmb@rasmb.org</a><br clear="none"><b>Subject:</b> Re: [RASMB] Centerpiece question</span><span style="font-size:12.0pt;"></span></div><div class="yiv1694068238MsoNormal"><span style="font-size:10.0pt;">John, in my view the advantages of the Stafford meniscus-matching design (available from Spin Analytical) over the synthetic boundary centerpieces are that the long capillary design inhibits diffusion between the two channels and any inadvertent fluid transfer during loading and handling. The drawback of that long channel though is that it is harder to get the transfer needed to match the heights (it takes more force)---I find I may need to go to 10-12 K rpm. For some of the virus samples we run that is >= to the run speed, so you have to do it very quickly to avoid significant sedimentation during the meniscus matching spin.</span></div><div class="yiv1694068238MsoNormal"><span style="font-size:10.0pt;"> </span></div><div class="yiv1694068238MsoNormal"><span style="font-size:10.0pt;">John</span></div><div class="yiv1694068238MsoNormal"><span style="font-size:10.0pt;"> </span></div><div><div style="border:none;border-top:solid #E1E1E1 1.0pt;padding:3.0pt 0in 0in 0in;"><div class="yiv1694068238MsoNormal"><b>From:</b> RASMB [<a rel="nofollow" shape="rect" ymailto="mailto:rasmb-bounces@list.rasmb.org" target="_blank" href="mailto:rasmb-bounces@list.rasmb.org">mailto:rasmb-bounces@list.rasmb.org</a>] <b>On Behalf Of </b>John Sumida<br clear="none"><b>Sent:</b> Thursday, December 18, 2014 9:45 AM<br clear="none"><b>To:</b> <a rel="nofollow" shape="rect" ymailto="mailto:rasmb@rasmb.org" target="_blank" href="mailto:rasmb@rasmb.org">rasmb@rasmb.org</a><br clear="none"><b>Subject:</b> [RASMB] Centerpiece question</div></div></div><div class="yiv1694068238MsoNormal"> </div><div class="yiv1694068238MsoNormal"><span style="font-size:12.0pt;">Dear RASMB,</span></div><div class="yiv1694068238MsoNormal"><span style="font-size:12.0pt;"> </span></div><div class="yiv1694068238MsoNormal"><span style="font-size:12.0pt;">For interference measurements where matching the sample and reference channel menisci is essential, is there a preference for either the meniscus matching centerpiece versus synthetic boundary centerpiece. We would be interested to hear about pros and cons.</span></div><div class="yiv1694068238MsoNormal"><span style="font-size:12.0pt;"> </span></div><div class="yiv1694068238MsoNormal"><span style="font-size:12.0pt;">Thank you and happy holidays.</span></div><div class="yiv1694068238MsoNormal"><span style="font-size:12.0pt;"> </span></div><div class="yiv1694068238MsoNormal"><span style="font-size:12.0pt;">Best regards,</span></div><div class="yiv1694068238MsoNormal"><span style="font-size:12.0pt;"> </span></div><div class="yiv1694068238MsoNormal"><span style="font-size:12.0pt;">John Sumida, Ph.D.</span></div><div class="yiv1694068238MsoNormal"><span style="font-size:12.0pt;"><a rel="nofollow" shape="rect" target="_blank" href="http://depts.washington.edu/cidb4bio/index.shtml"><span style="color:windowtext;">Analytical Biopharmacy Core Facility</span></a></span></div><div class="yiv1694068238MsoNormal"><span style="font-size:12.0pt;">University of Washington</span></div><div class="yiv1694068238MsoNormal"><span style="font-size:12.0pt;"><a rel="nofollow" shape="rect" target="_blank" href="http://www.moles.washington.edu/"><span style="color:windowtext;">Molecular Engineering & Sciences Institute, G22</span></a></span></div><div class="yiv1694068238MsoNormal"><span style="font-size:12.0pt;">3946 West Stevens Way NE</span></div><div class="yiv1694068238MsoNormal"><span style="font-size:12.0pt;">Seattle WA 98195-1653</span></div><div class="yiv1694068238MsoNormal"><span style=""> </span></div></div></div>_______________________________________________<br clear="none">RASMB mailing list<br clear="none"><a rel="nofollow" shape="rect" ymailto="mailto:RASMB@list.rasmb.org" target="_blank" href="mailto:RASMB@list.rasmb.org">RASMB@list.rasmb.org</a><br clear="none">http://list.rasmb.org/listinfo.cgi/rasmb-rasmb.org<br clear="none"><br clear="none"><div><br clear="none"></div>
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