[Histonet] H & E Troubleshooting

Wed Feb 24 08:21:12 CST 2016

"Another theory floating in the lab is because we bleach the stain buckets over the weekend maybe the bleach is saturating the bucket and slowly leaching out diluting the hematoxylin later in the day. (I would think this would be the case for all the slides not just the last few)"

I guess I need to give more details...the first few runs look "fine" per my supervisor. If we didn't rinse the buckets I would think the first few runs would look pale too...Leaching out bleach later in the day seems illogical to me. This is why I am leaning toward the xylene being saturated (but we didn't have half the cases that we normally do and it was fresh Monday morning when this occurred) Or the tap water which is hooked directly to our stainer and can vary from clean to yellow to tan within a day. I had this issue with the tap water when I was at another facility a few block from this one in the same city. I just need another voice of reason to show my supervisor.

Cassandra Davis
Histology Technician
Anatomical Pathology Laboratory
Saint Francis Healthcare
701 N. Clayton Street
Wilmington,DE 19805
Office:  302-575-8095
Email:  CDavis at che-east.org
www.saintfrancishealthcare.org
________________________________________
From: Podawiltz, Thomas [tpodawiltz at lrgh.org]
Sent: Tuesday, February 23, 2016 1:28 PM
To: Cassie P. Davis
Subject: RE: H & E Troubleshooting

Hi Cassandra,

Does the Eosin looked washed out. If so it may be your bluing. I had this issue a year or so ago and it turned out that the bluing was to strong and it was effecting the ph of the Eosin and giving it a washed out look. Since I had a gallon of the stuff, I just adjusted the time in bluing and my water rinse and the issue went away.

I will say I have also had the issue with bleached containers, but when that happened everything look light.

Tom

Tom Podawiltz HT (ASCP)
AP  Section Head
LRGHealthcare
603-524-3211 ext: 3220

-----Original Message-----
From: Cassie P. Davis via Histonet [mailto:histonet at lists.utsouthwestern.edu]
Sent: Tuesday, February 23, 2016 1:12 PM
To: histonet at lists.utsouthwestern.edu
Subject: [Histonet] H & E Troubleshooting

Hi Histo folks,

         We are having a fun time troubleshooting our H&E because the problem in not consistent...

Yesterday we poured fresh xylene, Hematoxylin , Eosin & bumped our alcohol since our ordered had not arrived yet. One of our doctor said our hematoxylin & eosin was readable but too light(he got 6 out of the 36 cases), the other (who got the 30 cases)said it was good.

We had half the number of slides going through staining than we normally do.

We don't let the last alcohol before xylene get pink from eosin.

Had it been a Thursday-Friday I would be concerned the xylene needed to be changed.

The Define and Bluing were made fresh.

My theory is the tap water could be off or the xylene needs to be changed.

Another theory floating in the lab is because we bleach the stain buckets over the weekend maybe the bleach is saturating the bucket and slowly leaching out diluting the hematoxylin later in the day. (I would think this would be the case for all the slides not just the last few)

Cassandra Davis
Histology Technician
Anatomical Pathology Laboratory
Saint Francis Healthcare
701 N. Clayton Street
Wilmington,DE 19805
Office:  302-575-8095
Email:  CDavis at che-east.org<mailto:n at che-east.org>
www.saintfrancishealthcare.org

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