[Histonet] DAKO LINK PTs

[Elizabeth Chlipala]

Tiana

We have never experienced any issue like that here we will use the HIER solutions in the PT links for up to three times within a period of 2 weeks.   We have had our PT link units validated and they are calibrated yearly.  Do you review the Target Retrieval Run Reports, we print and keep all of ours.  Our units are primarily programed to heat to 80C we put the slides in, warm up to 95C retrieve for 20 minutes cool down to 80C remove slides right after that.  I would think if you left slides in the retrieval solution for different times after they are completed you might see some changes in staining intensity, that why we try to be consistent and remove the slides as soon as they are completed.  We do use other retrieval times and temps on some occasions but what I stated is our standard protocol.   I hope this helps.  

There is one other thing, it think it is extremely important to clean the instrument as it is required, we have in our SOP's after 200 slides I don't know what Dako recommends but we actually clean more often typically after around 80 to 120 slides or so.  That will keep your probe nice and clean and decrease your variability, if you don't clean you can get build up in the probe and that is just going to cause inconsistent staining, its easy to set up a cleaning cycle at the end of the day.

There are also a lot of other factors that can affect staining consistency - tissue placement on the slide in one thing, we place our tissue in the same area on the slide, same number of drop zones, appropriate amount of reagents, cutting corners on amount of reagents will not lead to consistent high quality staining.    Section thickness can also lead to variation in staining intensity.  

Liz

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

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-----Original Message-----
From: Tiana Baskin via Histonet [mailto:histonet at lists.utsouthwestern.edu] 
Sent: Tuesday, July 14, 2015 8:42 AM
To: 'histonet at lists.utsouthwestern.edu'
Subject: [Histonet] DAKO LINK PTs

Hi Histonetters,

I have encountered a problem with some of my staining and I am struggling to pinpoint the root cause. I was wondering if any other DAKO Autostainer Link/PT users are experiencing the same oddities.

It seems like the first runs with fresh HIER PT solution is very typical of what we have optimized and the second batch of slides has more background and in some cases nuclear staining (especially in Actin (1A4) and pan CK (AE1/AE3)) when the target is cytoplasmic. We do not use the PT solution more than twice. What do others do? Have you seen similar things?

Tiana