Re: [Histonet] PAS-diastase staining

[Linda Prasad (SCHN)]

We have been making our own diastase solution for years now. Works extremely 
well. The diastase is very easy to make in house. Here is the protocol for 
making your own diastase

Principle:  
The enzyme solution is applied to one of two sections of the tissue (preferably 
consecutive sections) and then both are stained by the PAS method.  The 
presence and relative amount of glycogen in the sections can be determined by 
examining the extent of loss of staining in the enzyme treated section as 
compared with the untreated section.

This amylase reagent has a long shelf life and uses an incubation time of 10 
minutes at room temperature. It is suitable for formalin and Brazil’s fixed 
paraffin sections as well as air-dried and ethanol fixed frozen sections.

Controls:   Liver containing glycogen

Reagents:   
1.  Amylase Reagent
Warning: Harmful, contains azide – see MSDS
Alpha Amylase from Bacillus Subtilis (Sigma Cat No 10070,)  1g
Oxoid PBS Tablets (Cat No BR14a)1 tablet
Distilled water 100ml
Sodium Azide0.1g
This solution, once prepared is stored at 4oC when not in use. A recycled 
antibody dropper bottle (often used in commercial immunoperoxidase kits) is 
useful for storage and application.

2.  PAS Reagents (see PAS Stain)
Warning: Suspected Carcinogen – see MSDS
 

Procedure:

1.  Dewax and hydrate paraffin sections, hydrate frozen sections.
2.  For amylase digestion, place slides on a rack, cover sections with 
amylase solution and allow to incubate for 10 minutes at room temperature.
3.  Wash slides well in water.
4.  Place slides in 1% periodic acid 10 minutes.
5.  Wash slides well in water.
6.  Rinse slides in distilled water.
7.  Place in Schiff’s reagent 10 minutes.
8.  Rinse slides in distilled water and then wash slides in tap water 3 
minutes.
9.  Counterstain slides with haematoxylin, differentiate and blue.
10. Dehydrate, clear and mount.

Results: 
•   Glycogen is extracted and so loss of PAS positive staining will occur 
in the enzyme treated section.

•   Mucopolysaccharides are not extracted and so staining will be the same 
in both sections.

Reference:   
V-M. Mangan, V. Farago, M. Kelly, and A. F. Henwood (2002) " An Amylase Reagent 
with a Long Shelf Life for the Removal of Glycogen from Tissue Sections" J 
Histotechnol. 25(3): 153-4.


Linda Prasad | Senior Scientist | Histopathology
t: (02) 9845 3306 | f: (02) 9845 3318 | e: linda.pra...@health.nsw.gov.au | w: 
www.schn.health.nsw.gov.au
Cnr Hawkesbury Road and Hainsworth Street, Westmead, NSW Australia
Locked Bag 4001, Westmead 2145, NSW Australia

♲  Please consider the environment before printing this email.

-Original Message-
From: Roy, Lisa [mailto:ro...@labcorp.com] 
Sent: Saturday, 30 May 2015 1:37 AM
To: Rene J Buesa; Julio Benavides; histonet@lists.utsouthwestern.edu
Subject: Re: [Histonet] PAS-diastase staining

I have been using the spit method for years with great results.  Depar slide to 
water, add saliva directly to slide for 10 minutes, rinse and stain PAS as 
usual.  Also American MasterTech has diastase malt that works well.
Lisa

-Original Message-
From: Rene J Buesa [mailto:rjbu...@yahoo.com] 
Sent: Friday, May 29, 2015 11:17 AM
To: Julio Benavides; histonet@lists.utsouthwestern.edu
Subject: Re: [Histonet] PAS-diastase staining

Sigma porcine diastase is very good.A cheaper option is very disgusting but had 
been done for decades, just spit on the section but that will carry bacteria 
and, although I have seen doing it, I have never done it.René J.  


 On Friday, May 29, 2015 10:01 AM, Julio Benavides 
 wrote:
   

 Hi there,

I am trying to do a PAA-diastase in formalin-fixed embedded samples (ovine 
liver). I would be most grateful if someone could give me a hand in:

-Protocol (enzyme concentration, time of digestion...) for diastase digestion 
(I am assuming that PAS afterwards is the normal protocol for PAS) -A 
commercial source for diastase working in these samples. Is the porcine amylase 
from sigma any good? cheaper options?

As always,

thank you very much for all your comments

Cheers

Julio


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 On Friday, May 29, 2015 10:01 AM, Julio Benavides 
 wrote:
   

 Hi there,

I am trying to do a PAA-diastase in formalin-fixed embedded samples (ovine 
liver). I would be most grateful if someone could give me a hand in:

-Protocol (enzyme concentration, time of digestion...) for diastase digestion 
(I am assuming that PAS afterwards is the normal protocol for PAS) -A 
commercial source for diastase working in these samples. Is the porcine amylase 
from sigma any good? cheaper options?

As always,

Re: [Histonet] Plasma/thrombin clot for cell block

[Linda Prasad (SCHN)]

Hi Cheryl

This is the technique we use.

Thromboplastin Cell Block method

Principle:

The Thromboplastin cell block is very gentle on cells and allows a wide range 
of routine immunohistochemical stains to be done.
Plasma, treated with EDTA, is mixed with the specimen in the presence of 
thromboplastin and calcium ions to form a cell clot.
This method is not suitable for cells fixed in formalin or specimens of bile 
fluid (bile enzymes prevent clot formation)

Solutions:

1.  Normal Plasma, containing EDTA , use out of date plasma obtained from 
Blood Bank. Freeze in 2ml aliquots. This plasma has been tested for known 
viruses and is considered safe.

2.  Thromboplastin, use out of date thromboplastin obtained from 
Haematology.

3.  1% calcium chloride

4.  10% formalin

Method:

1.  Spin fluid using a centrifuge.
2.  Decant off supernatant
3.  Resuspend pellet in 3 drops of plasma.
4.  Add 3 drops of thromboplastin, mix
5.  Add 3 drops of Calcium Chloride, mix gently
6.  Allow to stand undisturbed for 15-20 minutes
7.  Add 5-8ml 10% formalin and allow to fix
8.  Place in a labelled tissue cassette and process as usual.

Reference: 
Furtado (1970) Stain Technol 45(1):19-23.

Thank you :)

Linda Prasad | Senior Scientist | Histopathology
t: (02) 9845 3306 | f: (02) 9845 3318 | e: linda.pra...@health.nsw.gov.au | w: 
www.schn.health.nsw.gov.au


Cnr Hawkesbury Road and Hainsworth Street, Westmead, NSW Australia
Locked Bag 4001, Westmead 2145, NSW Australia

♲  Please consider the environment before printing this email.

-Original Message-
From: Cheryl [mailto:tkngfl...@yahoo.com] 
Sent: Wednesday, 6 May 2015 2:19 AM
To: histonet@lists.utsouthwestern.edu
Subject: [Histonet] Plasma/thrombin clot for cell block

HI Guys-

I need procedures including products to create a plasma cell block for non-gyn 
cytology.

The one we have uses bovine thrombin and calcium chloride (dihydrate) but it's 
gotten corrupted and I can't find the original resources.

Help?

Cheryl
tkngfl...@yahoo.com

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RE: [Histonet] Orange peel for GMS

[Linda Prasad (SCHN)]

What about plain boiled rice. Leave if out for a few days. You will get a lot 
of mould growing. Rice should be easy to cut as well.

Linda Prasad | Senior Scientist | Histopathology
t: (02) 9845 3306 | f: (02) 9845 3318 | e: linda.pra...@health.nsw.gov.au | w: 
www.schn.health.nsw.gov.au

Cnr Hawkesbury Road and Hainsworth Street, Westmead, NSW Australia
Locked Bag 4001, Westmead 2145, NSW Australia
♲  Please consider the environment before printing this email.

-Original Message-
From: histonet-boun...@lists.utsouthwestern.edu 
[mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Piche, Jessica
Sent: Saturday, 18 April 2015 2:55 AM
To: 'Blazek, Linda'; Bitting, Angela K.; 'T Williams'; 
histonet@lists.utsouthwestern.edu
Subject: RE: [Histonet] Orange peel for GMS

Hahahaha I bet not Linda!!!

Jessica

-Original Message-
From: histonet-boun...@lists.utsouthwestern.edu 
[mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Blazek, Linda
Sent: Friday, April 17, 2015 12:05 PM
To: Bitting, Angela K.; Piche, Jessica; 'T Williams'; 
histonet@lists.utsouthwestern.edu
Subject: RE: [Histonet] Orange peel for GMS

Our blue cheese GMS would make you never want to eat it again!

Linda

-Original Message-
From: histonet-boun...@lists.utsouthwestern.edu 
[mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Bitting, Angela 
K.
Sent: Friday, April 17, 2015 11:21 AM
To: Piche, Jessica; 'T Williams'; histonet@lists.utsouthwestern.edu
Subject: RE: [Histonet] Orange peel for GMS

We just tried a moldy  strawberry. It was loaded with fungus!

-Original Message-
From: histonet-boun...@lists.utsouthwestern.edu 
[mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Piche, Jessica
Sent: Friday, April 17, 2015 7:11 AM
To: 'T Williams'; histonet@lists.utsouthwestern.edu
Subject: RE: [Histonet] Orange peel for GMS

I got a nice GMS control from some moldy chicken in my fridge. I am also trying 
the onion. I haven't stained the onion yet but it actually cut beautifully so I 
have high hopes. I let you know how it turns out. Have a nice weekend!

Jessica Piche, HT(ASCP)

-Original Message-
From: histonet-boun...@lists.utsouthwestern.edu 
[mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of T Williams
Sent: Thursday, April 16, 2015 9:57 PM
To: histonet@lists.utsouthwestern.edu
Subject: [Histonet] Orange peel for GMS

I tried using a moldy orange peel as a GMS control, but the staining did not 
work.  I ran it in a 10 hour processing protocol.  Should I try something 
different?  Any suggestions would be extremely helpful - we're desperate for a 
GMS control.

Thanks,
T. Williams, HTL (ASCP), QIHC
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RE: [Histonet] RE: ORO tissue falling off

[Linda Prasad (SCHN)]

I Know fatty tissue is such a pain to cut. Bryan Llewellyn gave some really 
good techniques. Im going to try them out myself :)

Linda Prasad | Senior Scientist | Histopathology
t: (02) 9845 3306 | f: (02) 9845 3318 | e: linda.pra...@health.nsw.gov.au | w: 
www.schn.health.nsw.gov.au


Cnr Hawkesbury Road and Hainsworth Street, Westmead, NSW Australia
Locked Bag 4001, Westmead 2145, NSW Australia

♲  Please consider the environment before printing this email.


-Original Message-
From: Caroline Miller [mailto:mi...@3scan.com] 
Sent: Thursday, 16 April 2015 10:44 AM
To: Linda Prasad (SCHN)
Cc: Jo-Ann Bader, Ms.; histonet@lists.utsouthwestern.edu
Subject: Re: [Histonet] RE: ORO tissue falling off

+1 to Linda, but I have found no difference on overnight vs multiple days.

Fatty liver is hard to do on all counts! It is tough enough sometimes to get a 
decent section on the slide.

Thanks for the other suggestions, certainly something I would try in the future

Yours
Caroline

Caroline Miller (mills)
Director of Histology
3Scan, Inc
415-2187297

> On Apr 15, 2015, at 5:07 PM, Linda Prasad (SCHN) 
>  wrote:
> 
> Usually with the fatty tissues, I pick them up on superfrost slides and let 
> it air dry  for 2-3 days at room temperature and then perform the ORO stains. 
> So far they seem to stay on.
> 
> Linda Prasad | Senior Scientist | Histopathology
> t: (02) 9845 3306 | f: (02) 9845 3318 | e: 
> linda.pra...@health.nsw.gov.au | w: www.schn.health.nsw.gov.au
> 
> Cnr Hawkesbury Road and Hainsworth Street, Westmead, NSW Australia 
> Locked Bag 4001, Westmead 2145, NSW Australia
> 
> ♲  Please consider the environment before printing this email.
> 
> -Original Message-
> From: histonet-boun...@lists.utsouthwestern.edu 
> [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Jo-Ann Bader, 
> Ms.
> Sent: Thursday, 16 April 2015 1:40 AM
> To: histonet@lists.utsouthwestern.edu
> Subject: [Histonet] ORO tissue falling off
> 
> We are having difficulty with a particulate set of very, very fatty 
> mouse livers.  The normal livers from this set stay on the slides the 
> fatty livers fall off.  We have used different types of charged slides 
> and we have even tried to drench the charged slides in Stay-On, dry 
> them and then put the frozen tissues on (despirate times call for 
> despirate measures).  No luck  Does anyone have any other ideas.  Help 
> Help
> 
> Jo-Ann  Bader
> Histology Coordinator
> Goodman Cancer Research Center
> 1600 Pine Ave. W,
> Room 312
> Montreal Quebec, H3A 1A3
> Email: jo-ann.ba...@mcgill.ca<mailto:jo-ann.ba...@mcgill.ca>
> Office Tel:  514-398-5647
> Lab:  Tel:  514-398-8270
> 
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> **
> *** This email and any files transmitted with it are 
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> whom they are addressed. If you are not the intended recipient, please delete 
> it and notify the sender.
> 
> Views expressed in this message and any attachments are those of the 
> individual sender, and are not necessarily the views of The Sydney Children's 
> Hospitals Network.
> 
> This note also confirms that this email message has been virus scanned and 
> although no computer viruses were detected, The Sydney Childrens Hospital's 
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RE: [Histonet] Bouin's, formic acid & and sirius red staining

[Linda Prasad (SCHN)]

Hi Tryrone
A Mass Trichrome stain will stain up the collagen in the diseased kidney. If 
the tissue has already been fixed in Bouin's Solution you can omit the picric 
acid step. Another good stain is the Curtis's Van Gieson Stain. Ive sent you 
the protocol for botht he stains. Hope it helps.   

Masson Trichrome

Principal
This method depends on the special action of phosphomolybdic acid when combined 
with red aniline dye. The acid lifts the colour first from the collagen and 
only later from the cytoplasm. The differentiation is stopped at the point 
where the Collagen only is colourless, and then another aniline dye (light 
green) is added to stain the collagen

Reagents:   

1.  Ponceau Acid Fuchsin 
Warning: Suspected Carcinogen – see MSDS
Acid fuchsin (Sigma Aldrich CI 42685) 0.1g
Ponceau de xylidene (CI 16150)  1g  
Distilled Water 99ml
When dissolved, add 1ml Glacial Acetic Acid

2.  Light Green
Warning: Suspected Carcinogen – see MSDS
Light green (CI 42095)  2g  
Glacial Acetic acid   2ml
Distilled water 98ml

3.  Weigert's Iron Haematoxylin or Celestine Blue/Haematoxylin

4.  1% Phosphomolybdic Acid
Phosphomolybdic acid5g
Distilled water 500ml
 

Procedure:  

1.  Bring sections to distilled water.  
2.  Celestine blue  10 minute.
3.  Wash well in water.
4.  Harris' haematoxylin10 minutes.
5.  Differentiate and blue  
6.  Ponceau acid fuchsin solution   10min
7.  Quick rinse in tap water
8.  1% Phosphomolybdic acid 5min
9.  Do not rinse in water
10. Light green solution2min 
11. Rinse excess stain rapidly in water.
12. Dehydrate, clear & mount

Results:  
Collagen, mucin green
Muscle, Fibrin  red
Nuclei  purple

Curtis's Van Gieson Stain

Principle:
This solution is based on the same formula as Van Gieson with the exception of 
the use of Ponceau S rather than acid fuschin. Glacial Acetic Acid is used 
rather than hydrochloric acid to sharpen the staining results.

Reagents: 

1.  1% aqueous Ponceau S 
Ponceau S (CI 27195)0.5g
Distilled water 50ml

2.  Curtis’s Van Gieson
1% aqueous Ponceau S10ml
Saturated aqueous picric acid   90ml
Glacial Acetic Acid 1.5ml

3.   Celestine Blue – Haematoxylin procedure

Procedure:
1.  Dewax and hydrate sections.
2.  Celestine blue 5 minute wash.
3.  Wash well in water.
4.  Harris' haematoxylin 5 minutes.
5.  Wash in water and blue (do not differentiate).
6.  Curtis Van Gieson stain 10 minutes (do not wash).
7.  Rinse in absolute alcohol 
8.  Dehydrate, clear and mount.

Results: 
CollagenRed
Muscle, other tissues   Yellow
Nuclei  Blue


Linda Prasad | Senior Scientist | Histopathology
t: (02) 9845 3306 | f: (02) 9845 3318 | e: linda.pra...@health.nsw.gov.au | w: 
www.schn.health.nsw.gov.au

Cnr Hawkesbury Road and Hainsworth Street, Westmead, NSW Australia
Locked Bag 4001, Westmead 2145, NSW Australia

♲  Please consider the environment before printing this email.


-Original Message-
From: histonet-boun...@lists.utsouthwestern.edu 
[mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Tyrone Genade
Sent: Thursday, 16 April 2015 7:10 AM
To: histonet
Subject: [Histonet] Bouin's, formic acid & and sirius red staining

Hello,

Some questions regarding Bouin's solution.

I was told, back when I was doing my PhD and new very little, that I should fix 
my fish in Bouin's as it will decalsify the bones. Well, Bouin's fixed fish 
were easier to cut than PFA fixed fish... but I read today that by adding 
formic acid the decalsification is better. (I must confess, that after Buoin's 
fixation I still had to soak the tissue face in some dilute nitric acid now and 
then...) Another reference said that the formaldehyde should be replaced with 
formic acid. So which is it: add formic acid or replace formaldehyde? And if 
the former, how much do you add?

Second question: a colleague and I want to stain for collagen in diseased 
kidneys. The fixative of choice for soft tissue is Bouin's... But the staining 
protocol called for a solution of picric acid and sirius red. Is the picric 
acid needed if I haven't washed the picric acid from the Bouin's fixation out 
of the tissue? I was told once that the picric acid was for contrast... Is

[Histonet] RE: ORO tissue falling off

[Linda Prasad (SCHN)]

Usually with the fatty tissues, I pick them up on superfrost slides and let it 
air dry  for 2-3 days at room temperature and then perform the ORO stains. So 
far they seem to stay on.

Linda Prasad | Senior Scientist | Histopathology
t: (02) 9845 3306 | f: (02) 9845 3318 | e: linda.pra...@health.nsw.gov.au | w: 
www.schn.health.nsw.gov.au

Cnr Hawkesbury Road and Hainsworth Street, Westmead, NSW Australia
Locked Bag 4001, Westmead 2145, NSW Australia

♲  Please consider the environment before printing this email.

-Original Message-
From: histonet-boun...@lists.utsouthwestern.edu 
[mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Jo-Ann Bader, 
Ms.
Sent: Thursday, 16 April 2015 1:40 AM
To: histonet@lists.utsouthwestern.edu
Subject: [Histonet] ORO tissue falling off

We are having difficulty with a particulate set of very, very fatty mouse 
livers.  The normal livers from this set stay on the slides the fatty livers 
fall off.  We have used different types of charged slides and we have even 
tried to drench the charged slides in Stay-On, dry them and then put the frozen 
tissues on (despirate times call for despirate measures).  No luck  Does anyone 
have any other ideas.  Help Help

Jo-Ann  Bader
Histology Coordinator
Goodman Cancer Research Center
1600 Pine Ave. W,
Room 312
Montreal Quebec, H3A 1A3
Email: jo-ann.ba...@mcgill.ca
Office Tel:  514-398-5647
Lab:  Tel:  514-398-8270

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RE: [Histonet] Allowable temperature range

[Linda Prasad (SCHN)]

If it's in formalin it can just stay at room temperature.

Linda Prasad | Senior Scientist | Histopathology
t: (02) 9845 3306 | f: (02) 9845 3318 | e: linda.pra...@health.nsw.gov.au | w: 
www.schn.health.nsw.gov.au


Cnr Hawkesbury Road and Hainsworth Street, Westmead, NSW Australia
Locked Bag 4001, Westmead 2145, NSW Australia
♲  Please consider the environment before printing this email.

-Original Message-
From: histonet-boun...@lists.utsouthwestern.edu 
[mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Tim H
Sent: Thursday, 2 April 2015 1:52 AM
To: histonet@lists.utsouthwestern.edu
Subject: [Histonet] Allowable temperature range


What is the allowable temperature range for a histology specimen after 
collection in formalin for shipping and storage?
 
Any ideas?
 Tim

  
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RE: [Histonet] Tol Blue

[Linda Prasad (SCHN)]

The mast cells are meant to turn purple. It’s a metachromatic stain. So the tol 
blue stains the tissue blue and the mast cells purple.

Mast cell granules  Purple
Acid Mucosubstance  Purple
Nuclei  Blue

Mast cells are rich in heparin. This allows them to be stained via acid 
mucopolysaccharide and metachromatic staining techniques. Churukian and Schenk 
developed this metachromatic dye-technique to reduce the excessive background 
staining that is common with metachromatic dye techniques.


Linda Prasad | Senior Scientist | Histopathology
t: (02) 9845 3306 | f: (02) 9845 3318 | e: linda.pra...@health.nsw.gov.au | w: 
www.schn.health.nsw.gov.au
Cnr Hawkesbury Road and Hainsworth Street, Westmead, NSW Australia
Locked Bag 4001, Westmead 2145, NSW Australia
♲  Please consider the environment before printing this email.

-Original Message-
From: histonet-boun...@lists.utsouthwestern.edu 
[mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Bryan Llewellyn
Sent: Wednesday, 25 March 2015 9:13 AM
To: histonet@lists.utsouthwestern.edu
Subject: Re: [Histonet] Tol Blue

Try the method at:

http://stainsfile.info/StainsFile/stain/cell/aldtolblue.htm

Bryan Llewellyn

Kimberly Marshall wrote:
> ?Hello my fellow Histo Techs.  Have a question I just know someone out there 
> can answer for me.
>
> In canine tissue, we are having problems with the Tol Blue for mast cell.  Am 
> experiencing metachromasia, or the mast cells turning purple not blue.  I 
> have attempted to decrease time, or add time, but its not helping.  My 
> pathologist says he has had this issue before.  So question is.  Could it be 
> the mast cell in a dog does not stain the same? Is there another stain that 
> may work?  Any help will be much appreciated.
>
>
> Thanks in Advance.
>
> Kimberly Marshall H.T.(ASCP)
>
>
>
>
>
>
> Kimberly Marshall H.T.(ASCP)
>
> Histology/Lab Supervisor
>
> Toll Free 1-800-426-2099
>
> Fax 801-584-5104
>
> PO Box 17580
>
> Salt Lake City, Utah 84107
>
> www.animalreferencepathology.com
>
>
>
> Advancing the art and science of veterinary medicine
>
>
>
> [cid:image001.jpg@01CF8F87.A0BD4830]
>
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[Histonet] RE: Mushrooms for GMS fungus control

[Linda Prasad (SCHN)]

I used strawberries for a fungal control. Worked really good.

Linda Prasad | Senior Scientist | Histopathology
t: (02) 9845 3306 | f: (02) 9845 3318 | e: linda.pra...@health.nsw.gov.au | w: 
www.schn.health.nsw.gov.au

Cnr Hawkesbury Road and Hainsworth Street, Westmead, NSW Australia
Locked Bag 4001, Westmead 2145, NSW Australia

♲  Please consider the environment before printing this email.

-Original Message-
From: histonet-boun...@lists.utsouthwestern.edu 
[mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Jeffrey Robinson
Sent: Saturday, 7 March 2015 4:16 AM
To: histonet@lists.utsouthwestern.edu
Subject: [Histonet] Mushrooms for GMS fungus control

How about mushrooms?  Has anyone had any success using mushrooms as a GMS 
fungus control?

Jeff Robinson, Senior Histotechnologist, Sierra Pathology Lab, Clovis, CA


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[Histonet] RE: PASD muscle stains

[Linda Prasad (SCHN)]

Usually we do the DiPAS stain on muscle frozen section. Air dried for 10 
minutes. Don't use any fixatives :)


Linda Prasad, MSc, BSc

Senior Scientist | Histopathology
t: 02 9845 3306 | f: 02 9845 3318 | e: linda.pra...@health.nsw.gov.au | w: 
www.schn.health.nsw.gov.au
m: 0425 314 267
the children's hospital at westmead
Cnr Hawkesbury Road and Hainsworth Street, Westmead
Locked Bag 4001, Westmead NSW 2145, AUSTRALIA



-Original Message-
From: histonet-boun...@lists.utsouthwestern.edu 
[mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Tiffany Passaro
Sent: Tuesday, 6 January 2015 10:23 AM
To: 'histonet@lists.utsouthwestern.edu'
Subject: [Histonet] PASD muscle stains

Greetings,


I am looking for fixatives that others are using in their labs 
for the PASD stain on fresh frozen muscle tissue. Currently we are fixing in 
10% NBF. Thanks in advance for any info on this.

Tiffany
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