<html><head><meta http-equiv="Content-Type" content="text/html charset=utf-8"></head><body style="word-wrap: break-word; -webkit-nbsp-mode: space; -webkit-line-break: after-white-space;" class=""><br class=""><div><blockquote type="cite" class=""><div class="">On Sep 14, 2017, at 8:35 PM, Masahide Kikkawa <<a href="mailto:mkikkawa@gmail.com" class="">mkikkawa@gmail.com</a>> wrote:</div><br class="Apple-interchange-newline"><div class=""><meta http-equiv="Content-Type" content="text/html; charset=utf-8" class=""><div style="word-wrap: break-word; -webkit-nbsp-mode: space; line-break: after-white-space;" class="">One of the advantage of one-sided blotting is the “filtering” effect of the holey carbon membrane for filamentous/tubular samples. <div class="">In this case, you can use lower concentrations of samples, because you can “concentrate” samples on the grid.<br class=""><div class="">
<span class="Apple-style-span" style="border-collapse: separate; font-family: 'Hiragino Kaku Gothic Pro'; font-style: normal; font-variant-ligatures: normal; font-variant-position: normal; font-variant-caps: normal; font-variant-numeric: normal; font-variant-alternates: normal; font-variant-east-asian: normal; font-weight: normal; letter-spacing: normal; line-height: normal; orphans: 2; text-indent: 0px; text-transform: none; white-space: normal; widows: 2; word-spacing: 0px; border-spacing: 0px; -webkit-text-decorations-in-effect: none; -webkit-text-stroke-width: 0px;"><div style="word-wrap: break-word; -webkit-nbsp-mode: space; -webkit-line-break: after-white-space; " class=""><span class="Apple-style-span" style="border-collapse: separate; font-family: 'Hiragino Kaku Gothic Pro'; font-size: inherit; font-style: normal; font-variant-ligatures: normal; font-variant-position: normal; font-variant-caps: normal; font-variant-numeric: normal; font-variant-alternates: normal; font-variant-east-asian: normal; 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-webkit-line-break: after-white-space; " class=""><span class="Apple-style-span" style="border-collapse: separate; font-family: 'Hiragino Kaku Gothic Pro'; font-size: 10px; font-style: normal; font-variant-ligatures: normal; font-variant-position: normal; font-variant-caps: normal; font-variant-numeric: normal; font-variant-alternates: normal; font-variant-east-asian: normal; font-weight: normal; letter-spacing: normal; line-height: normal; orphans: 2; text-indent: 0px; text-transform: none; white-space: normal; widows: 2; word-spacing: 0px; border-spacing: 0px; -webkit-text-decorations-in-effect: none; -webkit-text-stroke-width: 0px;"><div style="word-wrap: break-word; -webkit-nbsp-mode: space; -webkit-line-break: after-white-space; " class=""><span class="Apple-style-span" style="border-collapse: separate; font-family: 'Hiragino Kaku Gothic Pro'; font-size: 10px; font-style: normal; font-variant-ligatures: normal; font-variant-position: normal; font-variant-caps: normal; font-variant-numeric: normal; 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-webkit-text-stroke-width: 0px;"><div style="word-wrap: break-word; -webkit-nbsp-mode: space; -webkit-line-break: after-white-space; " class=""><div class=""><div style="word-wrap: break-word; -webkit-nbsp-mode: space; -webkit-line-break: after-white-space; " class=""><span class="Apple-style-span" style="border-collapse: separate; border-spacing: 0px; font-family: Osaka; font-size: 12px; font-style: normal; font-variant-ligatures: normal; font-variant-position: normal; font-variant-caps: normal; font-variant-numeric: normal; font-variant-alternates: normal; font-variant-east-asian: normal; font-weight: normal; letter-spacing: normal; line-height: normal; -webkit-text-decorations-in-effect: none; text-indent: 0px; text-transform: none; orphans: 2; white-space: normal; widows: 2; word-spacing: 0px;"><div style="word-wrap: break-word; -webkit-nbsp-mode: space; -webkit-line-break: after-white-space; " class=""><span class="Apple-style-span" style="border-collapse: separate; border-spacing: 0px; font-family: Osaka; font-size: 12px; font-style: normal; font-variant-ligatures: normal; font-variant-position: normal; font-variant-caps: normal; font-variant-numeric: normal; font-variant-alternates: normal; font-variant-east-asian: normal; font-weight: normal; letter-spacing: normal; line-height: normal; -webkit-text-decorations-in-effect: none; text-indent: 0px; text-transform: none; orphans: 2; white-space: normal; widows: 2; word-spacing: 0px;"><span class="Apple-style-span" style="border-collapse: separate; border-spacing: 0px; font-family: Osaka; font-size: 12px; font-style: normal; font-variant-ligatures: normal; font-variant-position: normal; font-variant-caps: normal; font-variant-numeric: normal; font-variant-alternates: normal; font-variant-east-asian: normal; font-weight: normal; letter-spacing: normal; line-height: normal; -webkit-text-decorations-in-effect: none; text-indent: 0px; text-transform: none; orphans: 2; white-space: normal; widows: 2; word-spacing: 0px;"><span class="Apple-style-span" style="border-collapse: separate; border-spacing: 0px; font-family: Osaka; font-size: 12px; font-style: normal; font-variant-ligatures: normal; font-variant-position: normal; font-variant-caps: normal; font-variant-numeric: normal; font-variant-alternates: normal; font-variant-east-asian: normal; font-weight: normal; letter-spacing: normal; line-height: normal; -webkit-text-decorations-in-effect: none; text-indent: 0px; text-transform: none; orphans: 2; white-space: normal; widows: 2; word-spacing: 0px;"><span class="Apple-style-span" style="border-collapse: separate; border-spacing: 0px; font-family: Osaka; font-size: 12px; font-style: normal; font-variant-ligatures: normal; font-variant-position: normal; font-variant-caps: normal; font-variant-numeric: normal; font-variant-alternates: normal; font-variant-east-asian: normal; font-weight: normal; letter-spacing: normal; line-height: normal; -webkit-text-decorations-in-effect: none; text-indent: 0px; text-transform: none; orphans: 2; white-space: normal; widows: 2; word-spacing: 0px;"><span class="Apple-style-span" style="border-collapse: separate; border-spacing: 0px; font-family: Osaka; font-size: 12px; font-style: normal; font-variant-ligatures: normal; font-variant-position: normal; font-variant-caps: normal; font-variant-numeric: normal; font-variant-alternates: normal; font-variant-east-asian: normal; font-weight: normal; letter-spacing: normal; line-height: normal; -webkit-text-decorations-in-effect: none; text-indent: 0px; text-transform: none; orphans: 2; white-space: normal; widows: 2; word-spacing: 0px;"><span class="Apple-style-span" style="border-collapse: separate; border-spacing: 0px; font-family: Osaka; font-size: 12px; font-style: normal; font-variant-ligatures: normal; font-variant-position: normal; font-variant-caps: normal; font-variant-numeric: normal; font-variant-alternates: normal; font-variant-east-asian: normal; font-weight: normal; letter-spacing: normal; line-height: normal; -webkit-text-decorations-in-effect: none; text-indent: 0px; text-transform: none; orphans: 2; white-space: normal; widows: 2; word-spacing: 0px;"><div class="">---------------------------------------------------------------</div><div class="">Masahide Kikkawa, M.D., Ph. D.</div><div class="">Professor</div><div class="">Department of Cell Biology & Anatomy</div><div class="">Graduate School of Medicine</div><div class="">The University of Tokyo</div><div class=""><a href="http://structure.m.u-tokyo.ac.jp/" class="">http://structure.m.u-tokyo.ac.jp/</a></div><div class="">tel: +81-3-5841-3338</div><div class=""><div class="">fax: +81-3-5841-3339</div><div class="">---------------------------------------------------------------</div></div></span></span></span></span></span></span></div></span></div></div></div></span></div></span></div></span></div></span></div></span></div></span></div><div class=""><blockquote type="cite" class=""><div class="">On Sep 15, 2017, at 12:31, Moni Banerjee <<a href="mailto:banerjee.moni@gmail.com" class="">banerjee.moni@gmail.com</a>> wrote:</div><div class=""><div dir="ltr" class=""><p class="MsoNormal"><span lang="EN-US" class="">Dear all,<span class=""></span></span></p><div class=""><span lang="EN-US" class=""> </span>I am wondering about the advantages (or
disadvantages) of one-sided vs two-sided blotting during sample freezing for
SPA. Do people have preferences for one or the other depending on the type of sample?
Does one-sided blotting lead to thicker ice in general? Any suggestion/comment
will be very welcome.</div><div class=""><span lang="EN-US" class=""> </span>Thanks,</div><div class=""><span lang="EN-US" class=""> </span>Manidipa</div><div class="gmail_signature"><div dir="ltr" class=""><div class="">------------------------<div class="">Dr. Manidipa Banerjee, PhD</div><div class="">Associate Professor</div><div class="">School of Biological Sciences</div><div class="">Indian Institute of Technology, Delhi</div><div class="">New Delhi 110016</div><div class="">India</div></div></div></div>
</div>
_______________________________________________<br class=""></div></blockquote></div><br class=""></div></div></div></blockquote></div>Dear Mandipa & Masahide,<div class=""><span class="Apple-tab-span" style="white-space:pre"> </span>One specimen I worked with had a much greater affinity for the blot paper than for the grid, so I had not only to try one-sided blotting, but I ended having to blot from the side of the grid to have the specimen stay on. This was a difficult process to get consistent ice thickness—mostly I failed—and it is a numbers game to get good enough specimens to get anything of value. One can get sufficiently thin ice from either one-sided or edge blotting, but it requires more skill. As usual, specimen preparation is the key to microscopy, and it is definitely specimen dependent.<br class=""><div class="">
<span class="Apple-style-span" style="border-collapse: separate; font-variant-ligatures: normal; font-variant-position: normal; font-variant-numeric: normal; font-variant-alternates: normal; font-variant-east-asian: normal; line-height: normal; border-spacing: 0px;"><div class=""><span class="Apple-tab-span" style="white-space: pre; "> </span>Yours,</div><div class=""><span class="Apple-tab-span" style="white-space: pre; "> </span>Bill</div><div class=""><br class=""></div></span><br class="Apple-interchange-newline">
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