[Histonet] FW: Leica Bond Max artifact

[Rene J Buesa]

Dear Dr. Rosen:
I see nothing wrong in your protocol but your photo 0950 = 09501 seems as if there are 2 superimposed sections, one with a lot of brown particles? Is this artifact you are referring to? Seems "too organized" to be an artifact but it is brown and not red.
These "non DAB" reagents sometimes develop a sort of precipitate even within their expiration date. Why have you decided to use this red reagent instead of DAB? That could be a better option.
Other than that I am also completely baffled by your problem.
René


 

    On Tuesday, October 9, 2018 1:15 PM, Linda Margraf via Histonet <histonet at lists.utsouthwestern.edu> wrote:
 

 Here is a message I am posting for Dr. Rosen:
Hello,

I am a longtime admirer of this list, and I am sure someone on here can help us out

We have an artifact on our immunohistochemistry (IHC) slides.

I will attempt to attach pictures using the upload tool. In case that doesn’t work, I am attaching a google drive link to some jpgs. The artifact is usually golden brown to red/pink in color, is clustered, appears both on top of the tissue and around it. If we do an aggressive rundown, it appears less red/brown and bluer.

We are using a Leica Bond MAX IHC system which is about a year old. We usually do two runs a day and 95% of the time we only use a red detection kit. We only see this on our red detection kit on our Leica bond. The brown is unaffected. The corresponding H&E slides are fine.
We started having this artifact after a month or two of production. 85% of the time we have some sort of artifact. Sometimes its perfect, sometimes it makes the tissue unreadable. We haven’t figured out any precipitating factors.

We have tried a number of things, but we still can’t get control of this.
Here is a brief overview of our protocols:
- Processor: Leica ASP300s tissue processor.
-Parrafin = Mercedes HWWX (dual purpose embedding and infiltration parrafin) melting point 57c – we tried switching, and it didn’t help.
- We only use distilled water in our baths
- Mercedes Starfrost charged slides
- Oven for 30 min at 62c
- We always use a cold red detection kit
-Sometimes our runs are delayed but not more than 4 hours
- Rundown is done in our stainer (standardized), and slides are coverslipped with tape.
- The bulk fluid containers in the bond are cleaned weekly and refrigerated between runs.
- The probe is cleaned every 300 slides.
- Mixing wells are cleaned out every two weeks and replaced often.
-Covertiles are well tended to.

We have tried to correct this problem, but we can't seem to fix it.
Any ideas?

https://drive.google.com/open?id=1wUttTweP7SYLRM5pkGOBr0MBzCMrtJSA<https://urldefense.proofpoint.com/v2/url?u=https-3A__drive.google.com_open-3Fid-3D1wUttTweP7SYLRM5pkGOBr0MBzCMrtJSA&d=DwMFaQ&c=fHUKEfYp8ZKZp-Z4zyr9bXWWb-JCctOum5ZlzbjkVjM&r=1o7iZWfSCXEgDesmDMuV2rPUZJJnqv0nrvyFk2x0v0A&m=i7Ca5SrRojLnKG1c5k_oFTuocpFcJZ4BUGjdsiw477Q&s=v-vNoy8KgrPg0AxG3-Oxmp7s0DKlExpBrpdNhGkUWQ8&e=>


Jason R. Rosen, D.O.
Dermatopathologist
Premier Dermatology Partners
4675 Linton Boulevard, Suite 203
Delray Beach, FL 33445
(p) 561-499-5341
(f) 561-499-5343
(e) JRosen at totalderms.com<mailto:JRosen at totalderms.com>


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