[3dem] negative staining Mycoplasma?

[Sarah Shahmoradian]

Hi David,

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).

Also I recall a paper (Asadi et al., 2017, *Micron*) 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.

Best wishes,
Sarah



--
Sarah H. Shahmoradian, PhD
Senior Scientist, Center for Cellular Imaging and NanoAnalytics (C-CINA),
Biozentrum, University of Basel
Mattenstrasse 26 | D-BSSE | WRO-1058 | CH-4058 Basel | Switzerland
sarah.shahmoradian at unibas.ch
Tel. +41 77 481 67 10

On Mon, Nov 9, 2020 at 9:44 PM Morgan, David Gene <dagmorga at indiana.edu>
wrote:

> Hi,
>
>
>     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.
>
>
> --
>     politics is more difficult than physics.
>                                              A. Einstein
>
>             David Gene Morgan
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