[Histonet] Sudanblack B on FFPET

[John Kiernan]

It's true that Sudan black B won't stain ordinary lipids (fat, phospholipids etc) that are absent from paraffin sections. The lipid in lipofuscin is bound to protein strongly enough to resist extraction during passage through all the solvents used in preparing paraffin sections. Churukian's adaptation of the Lillie & Ashburn method (link to a free download methods book in my recent Histonet email) is OK. It is one of the Biological Stain Commission's tests for certifying Sudan black B.
John Kiernan
London, Ontario
= = =
________________________________
From: AJ Cabral via Histonet <histonet at lists.utsouthwestern.edu>
Sent: March 28, 2023 4:12 PM
To: Tony Henwood <afhenwood at outlook.com>
Cc: Histonet at lists.utsouthwestern.edu <histonet at lists.utsouthwestern.edu>; Bob Richmond <rsrichmond at gmail.com>
Subject: Re: [Histonet] Sudanblack B on FFPET

Sudan Black staining won’t work on FFPET. The alcohols and xylenes used in
the tissue processing dissolves the lipids in the tissue. However, you can
used formalin fixed tissue as an alternative if no frozen section is
available. Rinse the tissue in distilled water for several minutes, pat
dry, freeze the tissue on OCT, cut frozen sections and stain in Sudan
black.

Have you considered looking into acid phosphatase staining for lipofucshin?
It is non specific but it can be demonstrated in muscle biopsy.

Cheers,
Joanna



On Tue, Mar 28, 2023 at 12:22 PM Tony Henwood via Histonet <
histonet at lists.utsouthwestern.edu> wrote:

> I would also let the saturated solution stand for a few days. Like Oil Red
> O, it probably needs time to “mature”. I would also use a frozen section of
> skin as a control.
>
> Regards,
>
> Tony Henwood
> Sydney, Australia
>
> From: Bob Richmond via Histonet<mailto:histonet at lists.utsouthwestern.edu>
> Sent: Wednesday, 29 March 2023 4:51 AM
> To: Histonet at lists.utsouthwestern.edu<mailto:
> histonet at lists.utsouthwestern.edu>
> Subject: Re: [Histonet] Sudanblack B on FFPET
>
> >
> > Gudrun Lang in Austria asks:
> >
>
> >>Has anyone experience with Sudanblack B on paraffin slides for staining
> [lipofuscin]? A doctor wants the demonstration of the lipoid content of
> foamy cells or granulocytes in lung. I've found protocols that have
> incubation-times from 10 minutes to over-night. - I've made a saturated
> Sudan black B-solution in 70% ethanol and tried it with10 min on liver
> without real success.<<
>
> The main thing you need to do is demonstrate that it isn't hemosiderin with
> an iron stain (Perls prussian blue reaction), and perhaps also that it
> isn't acid-fast. Lipofuscin can be identified an H & E staining, except for
> these considerations.
>
> Bob Richmond
> Maryville, Tennessee
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