I would also look at the deparaffinization of the slides.  Spotty staining
can be caused by incomplete removal of paraffin due to incomplete drying
and water under the tissue or water in the xylene (clearant) or alcohol
building up in the xylene (clearant). Check your reagents and make sure the
paraffin on dried slides is evenly melted over the slide.  Spotty areas of
incompletely dried or metled paraffin can indicate water under the tissue
or not drying long enough.  Pam Marcum


> [Original Message]
> From: louise renton 
> To: ; 
 > Date: 7/5/2002 3:42:12 AM
> Subject: Re: immunostaining
>
> Anissa,

> Could this be due to droplet formation on the tissue section?  Three
things 

> come to mind: incomplete covering of the section by reagents, drying of
the 

> reagents on the sections or incomplete dewaxing prior to staining. I
would 

> imagine that double dipping would cause, if anything, a problem
underneath 

> the section and not on top of it. You don't say what the patchiness looks 

> like.. if it is dark non specific staining then I suppose it could be due
to 

> "bubbles" in the section trapping reagents beneath the tissue. I have not 

> encountered this problem, only one due to poor covering of the section,

> 

> 

> Best regards

> Louise Renton

> Bone Research unit

> Johannesburg

> South Africa

> 

> 

> >From: Anissa Choi 

> >To: histonet@pathology.swmed.edu

> >Subject: immunostaining

> >Date: Fri, 05 Jul 2002 00:12:54 -0700

> >

> >

> >Lately we have problem of patchy staining with our immuno slides. we

> >run the immuno slides using Ventana immunostainer. I was assured by

> >Ventana'technical specialist that there is nothing wrong with the
machine.

> >It was suggested to me that the staining problem may be due to:

> >1. Poor quality of the "charged" slides

> >2. double dipping of "charged" slides when picking up section---slide

> >   is dipped in water and for some reason,gets dry before it is dipped

> >   into the water again for picking up section.

> >

> >I would think the sections will detach from the slides if there is

> >problem with the "charged "slides. I wonder how it can affect the

> >staining.

> >

> >Thanks in advance for your opinion.

> >

> >

> >Anissa Choi

> >

> >

> >

> >_________________________________________________________________

> >MSN Photos is the easiest way to share and print your photos:

> >http://photos.msn.com/support/worldwide.aspx

> >

> >

> 

> 

> 

> 

> _________________________________________________________________

> Send and receive Hotmail on your mobile device: http://mobile.msn.com

> 



--- Pamela Marcum
--- mucram11@earthlink.net
--- EarthLink: The #1 provider of the Real Internet.