PAS staining RE: [Histonet] Frozen tissue washing from slide

From:Gayle Callis

With frozen sections, a gentle rinse is advisable.  We cut frozen sections 
destined for routine stains and go directly to room temperature NBF - air 
drying is not really necessary for routine stains but is for 
immunostaiing.  Let your frozen sections fix longer than 10 minutes to 
ensure good adherance to slide and fixed proteins.

You can proceed with a PAS stain just as you do a paraffin section, at room 

Is the 10% acid glacial acetic or hydrochloric acid? You did not specify 
which one.  If you do acid/alcohol rinse, 0.5 to 1% HCl in 70%  for 
regressive or Harris hematoxylin , or if you use acetic acid, 4% in water 
is sufficient with progressive Gill type hematoxylin.

>-----Original Message-----
>[] On Behalf Of
>Sent: Tuesday, September 27, 2005 6:01 PM
>Subject: [Histonet] Frozen tissue washing from slide
>I'm hoping someone can give me some idea why frozen muscle sections
>suddenly start washing away from slide while trying to do PAS w/wo
>Other muscle panel stains do okay in staining process.....just the PAS
>w/wo is
>recently having problems.
>-We use superfrost plus slides
>-Sections are fixed in 10% formalin prior to stain
>-Most water rinses are DI.  Only one tap water rinse step right after
>amylase digest.
>-Does periodic acid have to be room temp prior to staining?
>-Could Shiffs reagent be too cold?
>-does temperature of water rinses affect adherance of sections to
>-We use a 10% acid water rinse after heme staining.  I recall if acid
>is made incorrectly, tissue sections could wash much like they do if
>water in H+E stain is too strong.
>Thanks for any thoughts and help.
>Histonet mailing list
>Histonet mailing list

Gayle Callis
Research Histopathology Supervisor
Veterinary Molecular Biology
Montana State University - Bozeman
PO Box 173610
Bozeman MT 59717-3610
406 994-6367
406 994-4303 (FAX)

Histonet mailing list

<< Previous Message | Next Message >>