[Histonet] Fading of H&E controls

Vickroy, James jvickroy at SpringfieldClinic.com
Thu Jul 21 09:09:05 CDT 2016

We are getting ready for our first CAP inspection in the new laboratory.    I was checking over the H&E controls that we do daily and made a discovery.   Several of the first slides we stained, over a year ago, are now washed out.  The eosin staining is probably worse than the hematoxylin however both have faded.    The H&E slides are in a file box that sits on the counter.   I next checked our surgical slide files and found that there were a very few slides from the same time period that have faded but most were still very good.

One of the weird things I have found in the controls is that some slides are as vibrant and brilliant as the day we stained them and some are almost completely faded with little color.   I am trouble shooting to figure out why the slides are fading.  I am trying to think of the variables  between the H&E control slides and the routine surgical slides.   The only thing that is different is that the routine surgical slides are filed in a rack and not exposed to the fluorescent light all day.  Is this possible?

Other thoughts:

If it is not the exposure to light than I am wondering if there is a problem with the staining protocol.   Another variable is that we use recycled clearing reagents.   On the stainer we use Fisher's Citrus Clearing agent and we recycle.  We process with Clearite III.  I know xylene is the best but do not want to use Xylene because of health issues.   I also wonder if there may still be some alcohol in the sections since some frozen sections fade when the clearing is not complete.  I do think the fading problem would be more evident in the routine filed slides if the problem was a staining protocol problem, but the recycled clearing agent is still on the table of suspects.
In the beginning we did not change the reagents as often as we do now but of course our volumes have increased also.

Please share with me your thoughts.

Jim Vickroy
Histology Manager
Springfield Clinic, Main Campus, East Building
1025 South 6th Street
Springfield, Illinois  62703
Office:  217-528-7541, Ext. 15121
Email:  jvickroy at SpringfieldClinic.com<mailto:jvickroy at SpringfieldClinic.com>

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