[Histonet] Histonet Digest, Vol 180, Issue 7

[Tyrone Genade]

Hello,

An update on my question:

On Fri, Nov 9, 2018 at 12:22 PM <histonet-request at lists.utsouthwestern.edu>
wrote:

>
> Message: 1
> Date: Thu, 8 Nov 2018 15:45:35 -0600
> From: Tyrone Genade <tgenade at gmail.com>
> To: histonet <histonet at lists.utsouthwestern.edu>
> Subject: [Histonet] counter stains for Sudan Black B
> Message-ID:
>         <
> CAEYEE3nxb7nWezRbL_UODTnTJtqiDhRyxkSdmgv4kvMv_pqpuw at mail.gmail.com>
> Content-Type: text/plain; charset="UTF-8"
>
> Hello,
>
> I need a counterstain for use together with Sudan Black B. I am using this
> method: https://www.ncbi.nlm.nih.gov/pubmed/27812872 which uses Nuclear
> Fast Red. I don't have nuclear fast red... In an updated protocol by these
> authors they use hematoxylin but in a previous trial I found it was too
> dark and make visualizing the Sudan Black signal too difficult. I'm using
> Sigma's Mayer's Hematoxylin solution. It is past its sell-by date and just
> won't blue nicely any more..
>
> I have available Methyl Green, Neutral Red and  Safranin  O. The former
> washes out in aqueous/glycerol mounting medium (required for the protocol).
> How well does Neutral Red and Safranin work with an aqueous mounting
> medium?
>

There were no replies but Neutral Red worked.

Does anyone know of a paper describing the cellular structures stained by
Neutral red? https://en.wikipedia.org/wiki/Neutral_red states that is
stains the Golgi apparatus and Nissl bodies and that living cells take up
the stain in lysosomes.
http://stainsfile.info/StainsFile/stain/nuclei/neutred.htm says it stains
the nucleus. I see a darker stained red halo around the nucleus and the
nucleus is stained reddish. It also seems to stain the aggresome, that is
it stains small structures juxtaposed to the nucleus. The Sudan Black seems
to overshadow the stain. Any speculation what is going on? Is the neutral
red staining structures that would also contain lipofuscin?

Thanks
-- 
Tyrone Genade
Orange City, Iowa
tel: (+1) 712 230 4101
http://tgenade.freeshell.org
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