[Histonet] GI biopsy H&E staining

[Jennifer MacDonald]

Use a Gill hematoxylin.

Sent from my iPad

On May 10, 2013, at 11:05 AM, "Annie T. Walton" <AWalton <@t> oima.org> wrote:

> Hello all,
>
> Our Medical Director would like the goblet cells on our GI biopsies to
stain a darker blue. I hand stain using the following protocol:
>
>
>  *   Xylene, 3 changes
3 minutes each
>  *   100% Alcohol, 2 changes
1 minute each
>  *   95% Alcohol
1 minute
>  *   Tap water
rinse until water runs off evenly
>  *   Harris Hematoxylin
5 minutes
>  *   Water wash
rinse until water runs clear
>  *   Clarifier 2
5 seconds
>  *   Running water
30 seconds
>  *   Bluing Solution
1 minute
>  *   Tap water
rinse well
>  *   95% Alcohol
30 seconds
>  *   Eosin Y
1 minute
>  *   100% Alcohol, 2 changes
30 seconds
>  *   100% Alcohol, 2 changes
1 minute each
>  *   Xylene, 3 changes
1 minute each
>
> I have recently increased the hematoxylin time from 4 to 5 minutes and
decreased the time in clarifier from 25 to 5 seconds so the hematoxylin
doesn't get washed out as much, but it is not good enough. I also switched
from 70% alcohol to 95% before eosin and increased the time in eosin from
45 seconds to 1 minute to provide a greater contrast in hopes that that
would work, but the goblet cells are still not staining dark enough. Any
suggestions would greatly be appreciated!
>
> Thanks!
>
> Annie Walton, HTL (ASCP)
> Histology Technician
> Overlake Internal Medicine Associates Histology Lab
> 425-974-9643
> awalton <@t> oima.org
>
>
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