Re: [Histonet] GMS Positive Fungal Tissue Needed

[Leonard Ringo via Histonet]

Hi Kelly 

We have blocks that you could have. We are currently looking for gram
negative, and AFB positive controls if you have any please let us know.
Please let us know if you have any of those and we can work something
out. 

Thanks, 

Leonard Ringo 

Macneal Hospital 

Anatomic Pathology 

3231 s Euclid ave. 

708. 783.7441 

leon...@rrclinicallab.com 

On 2018-05-31 03:54 PM, Pairan, Kelly via Histonet wrote:

> Good Afternoon,
> We are in need of some GMS positive fungus tissue for positive controls.  
> Would anyone out there happen to have some blocks we could have?  We would be 
> willing to swap if we had something you needed.  We would also cover the cost 
> of shipping.
> 
> Thanks,
> Kelly
> 
> Kelly Pairan,  HT (ASCP)CM, QIHC (ASCP)
> Specialist II-Anatomic Pathology
> Department of Pathology and Laboratory Medicine
> Email:  kelly.pai...@nationwidechildrens.org
> ph: 614-722-5414
> fx: 614-722-3033
> 
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[Histonet] GMS Positive Fungal Tissue Needed

[Pairan, Kelly via Histonet]

Good Afternoon,
We are in need of some GMS positive fungus tissue for positive controls.  Would 
anyone out there happen to have some blocks we could have?  We would be willing 
to swap if we had something you needed.  We would also cover the cost of 
shipping.

Thanks,
Kelly

Kelly Pairan,  HT (ASCP)CM, QIHC (ASCP)
Specialist II-Anatomic Pathology
Department of Pathology and Laboratory Medicine
Email:  kelly.pai...@nationwidechildrens.org
ph: 614-722-5414
fx: 614-722-3033

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[Histonet] FW: Ventana Optiview kit

[Lima, Teresa via Histonet]

Hello Histonetters,
Is anyone having issues with increased background staining of routine 
antibodies like keratins, CD markers and neuroendocrine markers using Ventana's 
antibodies with the Optiview kit? We are down to 4 minute antigen retrieval and 
primary antibody incubation times and still getting nonspecific staining. Our 
medical director thinks their prediluted antibodies are too concentrated for 
the kit. There are just a handful of their antibodies that are approved for 
this kit so there are no protocols in the package inserts. When we ran their 
breast markers with Optiview the positivity rate was incomparable to the 
results we get with UltraView and since the breast markers are not FDA cleared 
for OptiView we did not switch them over. It made our pathologists very 
apprehensive that with OptiView our Her2 validation cases became very positive 
and did not correlate with our UltraView stained slides and our FISH results. I 
look forward to your insights.
Thanks everyone!
Terri

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[Histonet] Ventana Optiview kit

[Lima, Teresa via Histonet]

Hello Histonetters,
Is anyone having issues with increased background staining of routine 
antibodies like keratins, CD markers and neuroendocrine markers using Ventana's 
antibodies with the Optiview kit? We are down to 4 minute antigen retrieval and 
primary antibody incubation times and still getting nonspecific staining. Our 
medical director thinks their prediluted antibodies are too concentrated for 
the kit. There are just a handful of their antibodies that are approved for 
this kit so there are no protocols in the package inserts. When we ran their 
breast markers with Optiview the positivity rate was incomparable to the 
results we get with UltraView and since the breast markers are not FDA cleared 
for OptiView we did not switch them over. It made our pathologists very 
apprehensive that with OptiView our Her2 validation cases  became very positive 
and did not correlate with our UltraView stained slides and our FISH results. I 
look forward to your insights.
Thanks everyone!
Terri

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the intended recipient, please notify me immediately as use of this information 
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[Histonet] Her2-ish

[Charles Riley via Histonet]

Does anyone run her2 ish on the Leica Bonds. If so who do you purchase your
antibody from?

-- 

Charles Riley BS  HT, HTL(ASCP)CM

Histopathology Coordinator/ Mohs
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[Histonet] who cut the frozen

[Cassie P. Davis via Histonet]

 Allison,
Here and the other three labs I worked at full-time, if the frozen arrived 
during "scheduled hisology hours", the histology techs cut them as long as it 
wouldn't accrue OT, after the techs left if the PA was still there, the PA 
would cut them , if the histology techs and PA left, then the pathologist would 
cut them.

Cassandra Davis
Histology Technician
AP Laboratory
302-575-8095
Email:  cda...@che-east.org



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Re: [Histonet] Masson's Trichrome Troubleshooting

[Mayer,Toysha N via Histonet]

Tasha,

Are you using the PolySci kit k037?  Did they change the formulation of the 
Bouins?  They do make one that has no picric acid, and doesn't last long.  If 
so, add a some saturated picric acid to the Bouins to see if that helps.
Are you using the Bouins again after microwaving?  If so, that may be your 
problem.  
Don't re-use the Phosphotungstic/phosphomolybdic acid, more than 2x.  It sets 
and diff's the red.

After the Anile blue, is the tissue blue?  If not, then it's the ppa/pma is the 
problem.  
Use the acetic acid more sparingly.  That diff's the blue some and can take 
more of it out.  I do it by sight.  Carson's says 3-5 min, but that is way too 
long.
We don't air dry, but that wouldn't take out the blue.  
Let me know if this helps.


Toysha N. Mayer D.H.Sc., MBA, HT(ASCP)
Instructor/Education Coordinator
HTL Program
MD Anderson School of Health Professions
713.563.3481
tnma...@mdanderson.org


--

Message: 2
Date: Wed, 30 May 2018 12:15:22 +
From: "Campbell, Tasha M." 
To: "histonet@lists.utsouthwestern.edu"

Subject: [Histonet] Masson's Trichrome Troubleshooting
Message-ID:

Content-Type: text/plain; charset="us-ascii"

Hello all,

I am having issues with my trichrome stain and I am about to lose my mind!  We 
just started doing it in house (although at my previous job I had done 
trichrome by hand for years so I am not a stranger to it. And I never had 
issues with it).  When I brought it in house at my current lab, I ordered the 
same kit that I was familiar with.  Its PolySci.  I did the stain about 5 or 6 
times and then all the sudden it quit working.  There was red where it should 
be blue.  And there was blue staining but it was all in the crypts.   I tried 
tweaking the stain a few times and nothing worked so I got a new kit.  The 
first 2 times I used the new kit, it worked perfect!  But after that it is back 
to doing the same thing again!  The collagen is not staining blue.  It is 
staining red.  Can anyone please tell me why this is happening?  I never had 
this issue before!  Thanks in advance!! See my protocol below.

1. Mordant in Bouin's solution, microwave 5 minutes, allow to stand 15 minutes.
 2. Wash in running tap water to remove the picric acid, 5 minutes.
 3. Weigert's Working Hematoxylin, 10 minutes.
 4. Blue in running tap water for 5 minutes, rinse in distilled water.
 5. Biebrich scarlet for 5 minutes.
 6. Rinse in distilled water.
 7. Phosphotungstic/phosphomolybdic acid for 10 minutes.
 8. Transfer directly into Aniline blue for 5 minutes.
 9. Rinse in distilled water.
10. 1% Acetic acid for 1 minute.
11. Quick rinse and air dry.
12. Coverslip




Tasha Campbell, B.S.,HTL(ASCP)
Frederick Gastroenterology Associates
310 W. 9th St.
Frederick, MD 21701
301-695-6800 ext. 144





--

Message: 5
Date: Wed, 30 May 2018 15:47:39 +0200
From: "Dr. Michael Gudo (Morphisto GmbH)" 
To: "Campbell, Tasha M." 
Cc: "histonet@lists.utsouthwestern.edu"

Subject: Re: [Histonet] Masson's Trichrome Troubleshooting
Message-ID: <5661355a-5f32-46eb-9f53-79fc9e494...@morphisto.de>
Content-Type: text/plain;   charset=utf-8

Dear Tasha,

well, I think you should modify your protocol:

(1) don?t use microwave
(2) wash in ethanol after Bouin and not in water (70 - 80 %)
(3) tap water as long as Weigert working solution
(4) Bieberich Scarlet is not the correct stain for Masson, you should use Acid 
fuchsine + Ponceau 2R + Azophloxine
(5) use a higher concentration of phosphortungstic acid, or / and least leave 
the sections much longer, up to 30 minutes in this step
(6) do not ?air dry?, but remove water with ethanol and ethanol with xylene, 
then you should get very nice results.

Well, and finally, if you allow I would like to offer you the Masson Kit / 
Masson Goldner Kit our company offers for quite brilliant stainings: 

https://webshop.morphisto.de/catalogsearch/result/?q=Kit+MASSON+trichrom 


Please do not hesitate to contact us, if you have any further questions.

With best regards
Michael




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Re: [Histonet] Masson's Trichrome Troubleshooting

[Gudrun Lang via Histonet]

Hi,
Check the staining after the PTA/PMA step. The acid should destain all the
tissue areas, where afterwards the anilinblue should bind. The acid replaces
the Biebrich scarlett in this areas and enables and enhances the binding of
anilinblue.
If the differentiation in insufficient, it may be the quality of the acid to
be the culprit. The pH should be about 2, if I remember correctly.

Check the temperature of the bouin after the microwave. If you cook the
tissue, it may also change the stainability. Microwave is not microwave, the
instrument in your old lab may have different characteristics.
A softer way is 60° for two hours in the oven.

Gudrun

-Ursprüngliche Nachricht-
Von: Campbell, Tasha M. via Histonet
[mailto:histonet@lists.utsouthwestern.edu] 
Gesendet: Mittwoch, 30. Mai 2018 14:15
An: histonet@lists.utsouthwestern.edu
Betreff: [Histonet] Masson's Trichrome Troubleshooting

Hello all,

I am having issues with my trichrome stain and I am about to lose my mind!
We just started doing it in house (although at my previous job I had done
trichrome by hand for years so I am not a stranger to it. And I never had
issues with it).  When I brought it in house at my current lab, I ordered
the same kit that I was familiar with.  Its PolySci.  I did the stain about
5 or 6 times and then all the sudden it quit working.  There was red where
it should be blue.  And there was blue staining but it was all in the
crypts.   I tried tweaking the stain a few times and nothing worked so I got
a new kit.  The first 2 times I used the new kit, it worked perfect!  But
after that it is back to doing the same thing again!  The collagen is not
staining blue.  It is staining red.  Can anyone please tell me why this is
happening?  I never had this issue before!  Thanks in advance!! See my
protocol below.

1. Mordant in Bouin's solution, microwave 5 minutes, allow to stand 15
minutes.
 2. Wash in running tap water to remove the picric acid, 5 minutes.
 3. Weigert's Working Hematoxylin, 10 minutes.
 4. Blue in running tap water for 5 minutes, rinse in distilled water.
 5. Biebrich scarlet for 5 minutes.
 6. Rinse in distilled water.
 7. Phosphotungstic/phosphomolybdic acid for 10 minutes.
 8. Transfer directly into Aniline blue for 5 minutes.
 9. Rinse in distilled water.
10. 1% Acetic acid for 1 minute.
11. Quick rinse and air dry.
12. Coverslip




Tasha Campbell, B.S.,HTL(ASCP)
Frederick Gastroenterology Associates
310 W. 9th St.
Frederick, MD 21701
301-695-6800 ext. 144

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