[Histonet] Tissue Washing

[Kathleen Roberts]

Emily,

The last time I had this problem, we switched to Ultrastick slides 
(ThermoFisher Scientific) and distilled water in the water bath.  I 
haven't had any problems since with H&E staining.

Kathleen Roberts
Principal Lab Technician
Neurotoxicology Labs
Dept of Pharmacology & Toxicology
Rutgers, the State University of NJ
41 B Gordon Rd
Piscataway, NJ 08854


Emily Butte wrote:

>Our lab has been struggling for a while now, with tissue washing off the
>slides. The problem was originally traced to our processor that was
>malfunctioning and having major carry-over issues, causing poor tissue
>processing. So after failed attempts to get the old machine fixed, we ended
>up getting a new microwave processor. The tissue (mainly prostate bx) comes
>out of the processor looking good. It cuts well, but after staining, some of
>the sections are no longer on the slides. The problem was quiet severe with
>the old processor and has decreased drastically, but has not entirely gone
>away with the microwave processor.
>
>Here's some info on our current procedure. This is a small physician office
>lab and we everything manual. We use plus slides and a water bath with no
>tissue adhesive added. The bath is between 42-45 degrees Celsius. After
>sectioning, the slide are put in an oven at 60 degrees for 20 minutes. After
>taking the slides out of the oven, we cool them to room temp then hand stain
>them in an H&E line.
>Xylene - 5 min
>Xylene - 2 min
>Xylene - 2 min
>100% Alcohol - 30 sec
>100% Alcohol - 30 sec
>100% Alcohol - 30 sec
>95% Alcohol - 30 sec
>Rinse in DiH2O
>Hematoxylin - 2 min
>Rinse in DiH2O
>0.50% Acid Alcohol - 2 dips
>Rinse in DiH2O
>Scott's Tap Water - 10 sec
>Rinse in DiH2O
>95% Alcohol - 30 sec
>Eosin - 30 sec
>95% Alcohol - 30 sec
>100% Alcohol - 30 secs
>100% Alcohol - 30 secs
>100% Alcohol - 30 secs
>Xylene - 30 sec
>Xylene 2 min
>Xylene - 2 min
>
>The slides are then cover slipped and of a 12 piece prostate case, at least
>a section or 2 will wash off of about 4 of the slides. Please pass along any
>ideas as to what the cause of this could be and how I can fix it. I'm fresh
>out of ideas and would really appreciate the advice of someone with more
>experience than myself.
>
>Thanks,
>  
>