Re: [Histonet] Porcessing FFPE tissue without alcohol??

2016-03-26 Thread Hobbs, Carl via Histonet 
Thanks, Liz.
If you look at fat all the time, using Osmium.you then are not sure if you 
use K-dichromate?
I am a tad confused

Alsowhy not trim the block too much?

Best wishes,

Carl
NB: Rene stated that I wouldn't be able to use any other fat stains...that's 
the point, Rene.
I don't need any other.
I commit to Osmium.

Yep...there are many variations for fat staining.
Imho...most are histochemical mythology.
Osmium "stains" fat...histologically.
As do  the conventional fat-soluble dyes when using frozen sections.
I would only listen to alternatives/disagreements from JKiernan.

I am still waiting for the next "generation" Histo person.
Cook, Kiernan.who are the other Seminals??

Enquiringly

Carl


From: Elizabeth Chlipala 
Sent: 26 March 2016 15:58
To: Joanna; Rene J Buesa
Cc: Hobbs, Carl; histonet
Subject: RE: [Histonet] Porcessing FFPE tissue without alcohol??

We use osmium post fixation to look at fat all of the time in mouse liver, 
nerve and muscle samples.  It works well, sample size needs to be thin, samples 
are friable and can crack easily.  We use a specific procedure for this it 
includes potassium dichromate I think, I'm at home but on Monday I can send the 
reference.  One more thing don't trim into the block too much.

Liz

Elizabeth A. Chlipala, BS, HTL(ASCP)QIHC
Premier Laboratory, LLC
PO Box 18592
Boulder, CO 80308
(303) 682-3949 office
(303) 881-0763 cell
(303) 682-9060 fax
l...@premierlab.com

Ship to address:

Premier Laboratory, LLC
1567 Skyway Drive, Unit E
Longmont, CO 80504

From: Joanna via Histonet [histonet@lists.utsouthwestern.edu]
Sent: Saturday, March 26, 2016 9:20 AM
To: Rene J Buesa
Cc: Hobbs, Carl; histonet
Subject: Re: [Histonet] Porcessing FFPE tissue without alcohol??

How about Sudan Black stain?

> On Mar 26, 2016, at 4:32 AM, Rene J Buesa via Histonet 
>  wrote:
>
> The only problem I see is that the fat will be preserved, as you wrote, as a 
> black osmium oxidate but you will not be able to use any "standard" fat 
> stain; otherwise it will work.René
>
>On Friday, March 25, 2016 2:41 PM, "Hobbs, Carl via Histonet" 
>  wrote:
>
>
>
>
> Fix the tissue in Formalin, wash well in dw, then place very small pieces 
> into Osmium tetroxide solution ( std soln for TEM post-fixation)
> Processing to Pwax as usual.
> Basically, you will see lipids as black ( oxidised osmium)
> That's the only way to demonstrate solvent- soluble lipids, using Pwax 
> processing.
> Sure, there are caveats but, in the main...it will be Ok, imho.
> I invite comments as I may be doing exactly this very soon, to count 
> myelinated nerve fibres in a sciatic nerve.
>
>
>
>
>
> Carl Hobbs FIBMS
> Histology and Imaging Manager
> Wolfson CARD
> Guys Campus, London Bridge
> Kings College London
> London
> SE1 1UL
>
> 020 7848 6813
> ___
> Histonet mailing list
> Histonet@lists.utsouthwestern.edu
> http://lists.utsouthwestern.edu/mailman/listinfo/histonet
>
>
> ___
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[Histonet] Processing FFPE tissue without alcohol

2016-03-26 Thread Wanda Shotsberger Gray via Histonet 


While the tissue will still go through alcohol, have you considered 
preserving the fat with osmium tetroxide prior to routine processing? This 
turns the fat black, but it is retained in the tissue.



Hi Histonet, is there a way to process tissue for paraffin embedding 
without using alcohol? One of the labs that send their processing to us 
will be doing a study examining fat. I told them if they want to look at 
the fat they will have to cut frozen sections but I'm being ask again about 
processing without the alcohol. So I said I would ask around.


Please let me know what you think.

Thank you for your help!

E-van



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Re: [Histonet] Porcessing FFPE tissue without alcohol??

2016-03-26 Thread Elizabeth Chlipala via Histonet 
We use osmium post fixation to look at fat all of the time in mouse liver, 
nerve and muscle samples.  It works well, sample size needs to be thin, samples 
are friable and can crack easily.  We use a specific procedure for this it 
includes potassium dichromate I think, I'm at home but on Monday I can send the 
reference.  One more thing don't trim into the block too much.

Liz

Elizabeth A. Chlipala, BS, HTL(ASCP)QIHC
Premier Laboratory, LLC
PO Box 18592
Boulder, CO 80308
(303) 682-3949 office
(303) 881-0763 cell
(303) 682-9060 fax
l...@premierlab.com

Ship to address:

Premier Laboratory, LLC
1567 Skyway Drive, Unit E
Longmont, CO 80504

From: Joanna via Histonet [histonet@lists.utsouthwestern.edu]
Sent: Saturday, March 26, 2016 9:20 AM
To: Rene J Buesa
Cc: Hobbs, Carl; histonet
Subject: Re: [Histonet] Porcessing FFPE tissue without alcohol??

How about Sudan Black stain?

> On Mar 26, 2016, at 4:32 AM, Rene J Buesa via Histonet 
>  wrote:
>
> The only problem I see is that the fat will be preserved, as you wrote, as a 
> black osmium oxidate but you will not be able to use any "standard" fat 
> stain; otherwise it will work.René
>
>On Friday, March 25, 2016 2:41 PM, "Hobbs, Carl via Histonet" 
>  wrote:
>
>
>
>
> Fix the tissue in Formalin, wash well in dw, then place very small pieces 
> into Osmium tetroxide solution ( std soln for TEM post-fixation)
> Processing to Pwax as usual.
> Basically, you will see lipids as black ( oxidised osmium)
> That's the only way to demonstrate solvent- soluble lipids, using Pwax 
> processing.
> Sure, there are caveats but, in the main...it will be Ok, imho.
> I invite comments as I may be doing exactly this very soon, to count 
> myelinated nerve fibres in a sciatic nerve.
>
>
>
>
>
> Carl Hobbs FIBMS
> Histology and Imaging Manager
> Wolfson CARD
> Guys Campus, London Bridge
> Kings College London
> London
> SE1 1UL
>
> 020 7848 6813
> ___
> Histonet mailing list
> Histonet@lists.utsouthwestern.edu
> http://lists.utsouthwestern.edu/mailman/listinfo/histonet
>
>
> ___
> Histonet mailing list
> Histonet@lists.utsouthwestern.edu
> http://lists.utsouthwestern.edu/mailman/listinfo/histonet

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Re: [Histonet] Porcessing FFPE tissue without alcohol??

2016-03-26 Thread Rene J Buesa via Histonet 
Sudan Black reacts only with protein-combined fats.René 

On Saturday, March 26, 2016 11:20 AM, Joanna  wrote:
 

 How about Sudan Black stain? 

> On Mar 26, 2016, at 4:32 AM, Rene J Buesa via Histonet 
>  wrote:
> 
> The only problem I see is that the fat will be preserved, as you wrote, as a 
> black osmium oxidate but you will not be able to use any "standard" fat 
> stain; otherwise it will work.René 
> 
>    On Friday, March 25, 2016 2:41 PM, "Hobbs, Carl via Histonet" 
> wrote:
> 
> 
> 
> 
> Fix the tissue in Formalin, wash well in dw, then place very small pieces 
> into Osmium tetroxide solution ( std soln for TEM post-fixation)
> Processing to Pwax as usual.
> Basically, you will see lipids as black ( oxidised osmium)
> That's the only way to demonstrate solvent- soluble lipids, using Pwax 
> processing.
> Sure, there are caveats but, in the main...it will be Ok, imho.
> I invite comments as I may be doing exactly this very soon, to count 
> myelinated nerve fibres in a sciatic nerve.
> 
> 
> 
>  
> 
> Carl Hobbs FIBMS 
> Histology and Imaging Manager 
> Wolfson CARD 
> Guys Campus, London Bridge  
> Kings College London 
> London 
> SE1 1UL 
>  
> 020 7848 6813    
> ___
> Histonet mailing list
> Histonet@lists.utsouthwestern.edu
> http://lists.utsouthwestern.edu/mailman/listinfo/histonet
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> 
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Re: [Histonet] Porcessing FFPE tissue without alcohol??

2016-03-26 Thread Joanna via Histonet 
How about Sudan Black stain? 

> On Mar 26, 2016, at 4:32 AM, Rene J Buesa via Histonet 
>  wrote:
> 
> The only problem I see is that the fat will be preserved, as you wrote, as a 
> black osmium oxidate but you will not be able to use any "standard" fat 
> stain; otherwise it will work.René 
> 
>On Friday, March 25, 2016 2:41 PM, "Hobbs, Carl via Histonet" 
>  wrote:
> 
> 
> 
> 
> Fix the tissue in Formalin, wash well in dw, then place very small pieces 
> into Osmium tetroxide solution ( std soln for TEM post-fixation)
> Processing to Pwax as usual.
> Basically, you will see lipids as black ( oxidised osmium)
> That's the only way to demonstrate solvent- soluble lipids, using Pwax 
> processing.
> Sure, there are caveats but, in the main...it will be Ok, imho.
> I invite comments as I may be doing exactly this very soon, to count 
> myelinated nerve fibres in a sciatic nerve.
> 
> 
> 
>   
> 
> Carl Hobbs FIBMS 
> Histology and Imaging Manager 
> Wolfson CARD 
> Guys Campus, London Bridge  
> Kings College London 
> London 
> SE1 1UL 
>   
> 020 7848 6813
> ___
> Histonet mailing list
> Histonet@lists.utsouthwestern.edu
> http://lists.utsouthwestern.edu/mailman/listinfo/histonet
> 
> 
> ___
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Re: [Histonet] Porcessing FFPE tissue without alcohol??

2016-03-26 Thread Rene J Buesa via Histonet 
The only problem I see is that the fat will be preserved, as you wrote, as a 
black osmium oxidate but you will not be able to use any "standard" fat stain; 
otherwise it will work.René 

On Friday, March 25, 2016 2:41 PM, "Hobbs, Carl via Histonet" 
 wrote:
 

 

Fix the tissue in Formalin, wash well in dw, then place very small pieces into 
Osmium tetroxide solution ( std soln for TEM post-fixation)
Processing to Pwax as usual.
Basically, you will see lipids as black ( oxidised osmium)
That's the only way to demonstrate solvent- soluble lipids, using Pwax 
processing.
Sure, there are caveats but, in the main...it will be Ok, imho.
I invite comments as I may be doing exactly this very soon, to count myelinated 
nerve fibres in a sciatic nerve.



  
 
Carl Hobbs FIBMS 
Histology and Imaging Manager 
Wolfson CARD 
Guys Campus, London Bridge  
Kings College London 
London 
SE1 1UL 
  
020 7848 6813    
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