[Histonet] RE: Pankeratin- Trouble with Mast Cell Staining

[HERRINGTON, SHEILA]

We found that if using the stronger proteases/enzymes for retrievals, there tended to be more mast cell staining seen.  


Sheila Herrington
Technical Lead - Histopathology and Immunohistochemistry
Kelowna General Hospital
2268 Pandosy Street, Kelowna, B.C. V1Y 1T2
250-862-4300 ext 7587 or 7510
Sheila.herrington <@t> interiorhealth.ca



-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu [mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Daniel Gonzalez
Sent: Tuesday, May 20, 2014 1:33 PM
To: 'Miller, Suzie'; histonet <@t> lists.utsouthwestern.edu
Subject: [Histonet] RE: Pankeratin- Trouble with Mast Cell Staining

Posting for my colleague, Jeff Gordon, because his post did not go through successfully:

"Suzie, your histonet post was forwarded to me by a colleague, and I thought I could help with your mast cell staining dilemma with pan-keratin.  University of Pennsylvania published an article a while back about "undesirable" reactivity of cytokeratins that focused primarily on sentinel lymph node testing.  In their study they found that when cytokeratin markers OTHER than strictly AE1/AE3 (with no other components) were used, other non-epithelial elements were being labeled that they deemed "undesirable."  This entire article can be found here for your's and your pathologists' reference:  http://www.archivesofpathology.org/doi/pdf/10.1043/0003-9985(2000)124%3C1310%3AUCIONN%3E2.0.CO%3B2  

Based on these findings, and because another person in the thread also indicated that they may be dealing with the same issue, cervical tissue that was rich in mast cells was stained on a Ventana Ultra with cytokeratin AE1/AE3 ONLY as well as with cytokeratin OSCAR (which is another pan-keratin but with a lesser sensitivity to squamous epithelium than AE1/AE3) with CC1 mild heat retrieval and 16 minute heated primary incubation and Ultraview detection.  To confirm the presence of mast cells in the tissue, both CD117 and tryptase were also stained on the same tissue with the same protocols.  The results showed that though there was an abundance of mast cells present labeled by tryptase and CD117, there was no detectable staining of mast cells with either cytokeratin AE1/AE3 or with cytokeratin OSCAR, while the epithelial portion of the tissue stained with both cytokeratins (though the OSCAR was noticeably weaker because of the lower sensitivity to squamous epithelium).  

The stains can be seen here:  http://www.cellmarque.com/cmc/AO_mastcellreactivity.php  

Hopefully this helps.  If you have any questions, please contact us at your convenience."

Thank you

-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu [mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Miller, Suzie
Sent: Tuesday, May 06, 2014 8:57 AM
To: histonet <@t> lists.utsouthwestern.edu
Subject: [Histonet] Pankeratin- Trouble with Mast Cell Staining

Hello Everyone,

If you have a Ventana Ultra and use (Ventana# 760-2595) Pankeratin AE1/AE3/PCK29, would you consider sharing your staining protocol?

Our Pathologists are displeased with the amount of mast cell staining with our current protocol so we are looking to explore other protocols or suggestions.

Thank you for your assistance,
Suzie

Suzie Miller, MLT ASCP
Senior Histotech
Mercy Health System of Maine Laboratory
millers <@t> emhs.org<mailto:millers <@t> emhs.org>

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