<div dir="ltr"><div dir="ltr"><div class="gmail_default" style="font-family:arial,helvetica,sans-serif;font-size:small;color:#000000">Hi David,</div><div class="gmail_default" style="font-family:arial,helvetica,sans-serif;font-size:small;color:#000000"><br></div><div class="gmail_default" style="font-family:arial,helvetica,sans-serif;font-size:small;color:#000000">Since Mycoplasma lack a cell wall unlike other bacteria, you may have better luck with negative staining in treating them like liposomes. For this, you could try phosphotungstic acid (PTA), i.e neutral PTA, 1% or 2% in water at pH 7 (protect from light and keep at 4ºC). Prior to using the PTA, you may want to filter/sonicate it to remove any aggregates (i.e. 13,800 × g for 10 min).</div><div class="gmail_default" style="font-family:arial,helvetica,sans-serif;font-size:small;color:#000000"><br></div><div class="gmail_default" style="font-family:arial,helvetica,sans-serif;font-size:small;color:#000000">Also I recall a paper (Asadi et al., 2017, <i>Micron</i>) that used embedment in methylcellulose films containing a mix of both uranyl acetate and phosphotungstic acid as a contrasting agent to stabilize and enhance contrast of these "floppy" type of lipid rich structures. You may find it useful for applying to your Mycoplasma samples.</div><div class="gmail_default" style="font-family:arial,helvetica,sans-serif;font-size:small;color:#000000"><br></div><div class="gmail_default" style="font-family:arial,helvetica,sans-serif;font-size:small;color:#000000">Best wishes,</div><div class="gmail_default" style="font-family:arial,helvetica,sans-serif;font-size:small;color:#000000">Sarah</div><div class="gmail_default" style="font-family:arial,helvetica,sans-serif;font-size:small;color:#000000"><br></div><div class="gmail_default" style="font-family:arial,helvetica,sans-serif;font-size:small;color:#000000"><br></div><div class="gmail_default" style="font-family:arial,helvetica,sans-serif;font-size:small;color:#000000"><br></div><div class="gmail_default" style="font-family:arial,helvetica,sans-serif;font-size:small;color:#000000">--</div><div class="gmail_default" style="font-family:arial,helvetica,sans-serif;font-size:small;color:#000000">Sarah H. Shahmoradian, PhD</div><div class="gmail_default" style="font-family:arial,helvetica,sans-serif;font-size:small;color:#000000">Senior Scientist, Center for Cellular Imaging and NanoAnalytics (C-CINA), Biozentrum, University of Basel<br>Mattenstrasse 26 | D-BSSE | WRO-1058 | CH-4058 Basel | Switzerland<br><a href="mailto:sarah.shahmoradian@unibas.ch" target="_blank">sarah.shahmoradian@unibas.ch</a><br>Tel. +41 77 481 67 10<br></div></div><br><div class="gmail_quote"><div dir="ltr" class="gmail_attr">On Mon, Nov 9, 2020 at 9:44 PM Morgan, David Gene <<a href="mailto:dagmorga@indiana.edu" target="_blank">dagmorga@indiana.edu</a>> wrote:<br></div><blockquote class="gmail_quote" style="margin:0px 0px 0px 0.8ex;border-left:1px solid rgb(204,204,204);padding-left:1ex">
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<p>Hi,</p>
<p><br>
</p>
<p> By some chance, does anyone have experience negatively staining Mycoplasma cells grown in culture? I tried what I would normally do with a bacterial culture and was rather disappointed, so any suggestions and tricks would be welcome. Thanks in advance.<br>
</p>
<p><br>
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