[Histonet] granules after X-gal staining

[Rachel, Rivka (NIH/NEI) [E]]

We used to see this all the time and couldn't figure out the problem.  The histologist even tried filtering all the solutions used for perfusion, staining solutions and many other things but never could determine the cause.  Any suggestions?


-----Original Message-----
From: koellingr <@t> comcast.net [mailto:koellingr <@t> comcast.net]
Sent: Fri 4/4/2008 12:16 AM
To: Yves Heremans; histonet <@t> lists.utsouthwestern.edu
Subject: Re: [Histonet] granules after X-gal staining
 
I haven't seen any replies and I am scientifically curious.  The beta-galactosidase is simply the enzyme that splits x-gal to eventually produce that classical blue chromogen deposit.  The acinar cells of pancreas are laden with known ..ase's (lipoxygenase, proteases, amylase, lipase, elastase, tryptase, etc, etc ase's) and probably unidentified ones.  Is it possible that some promiscuous enzyme is substituting enzymatically for beta-galactosidase to get your staining of tiny round blue granules in cytoplasm.  If you are working in frozens, your enzymes could all be very active.  Have looked at lots of x-gal staining but never in pancreas.  Have you stained a normal, not b-gal expressing, mouse pancreas? Am curious and hope someone has done this.

Ray Koelling
PhenoPath Labs
Seattle, WA

-------------- Original message -------------- 
From: Yves Heremans <Yves.Heremans <@t> vub.ac.be> 

> Dear Histonetters, 
> 
> Does anyone know why I am getting granules (tiny, round blue granules 
> in the cytoplasm) after X-gal staining on frozen sections of mouse 
> pancreas ? 
> 
> Regards, 
> 
> Yves 
> 
> 
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