Re: How long are subbed slides good?

From:Melissa Jensen

We haven't had any problems with special stains,immunos,or anything while
using stayon.We put all immuno sections on  positive charged slides.
----- Original Message -----
From: "rueggp" 
To: "Melissa Jensen" 
Cc: ; "Cathy Gorrie" 
Sent: Friday, April 12, 2002 12:53 PM
Subject: Re: How long are subbed slides good?


> i thought things in the water bath like (Stay ON, if it goes in the WB)
negated
> the charge on subbed slides????
> patsy
>
> Melissa Jensen wrote:
>
> > Maybe using gelatin in your bath water is the problem.I worked in a
histo
> > lab prior to the one I am at.We only used gelatin there..Had trouble
keeping
> > tissue on slides during gms procedure etc.I was introduced to Stay on at
my
> > current facility.It is great! Nothing falls off..no curling,You don't
even
> > have to let slides cool before putting them in the first xylene....There
is
> > another product called Halt.We tried it..but it reduced the surface
tension
> > of the water so much..All tissue no matter what curled...didn't adhere
to
> > the slide......We use subbed slides..they all stay in tact..
> > Hope this helps!
> > ----- Original Message -----
> > From: "Cathy Gorrie" 
> > To: 
> > Sent: Thursday, April 11, 2002 7:43 PM
> > Subject: Re: How long are subbed slides good?
> >
> > > We regularly cut 30-40 µm rat brain sections, place on subbed slides
> > > (0.8% gelatin with 0.05% chromium potassium sulphate) and air dry
> > > overnight before staining. Normally this is absolutely fine,and the
> > > age of subbed slides has varied from a few weeks to very old.
> > >
> > > Recently however, we have been having trouble with the same problems
> > > you are experiencing, BUT only on one series and one type of
> > > staining, rather than just any liquid. ie sections have been cut and
> > > placed on slides in series of two, one stays on through the stain
> > > over 4 days but the Cresyl Violet (10 mins) stained sections are
> > > lifting. I don't think it is the age of the subbing therefore, but
> > > possibly is due to the quality of subbing or staining solutions.
> > > There was some discussion on this list a while ago about alkaline
> > > solutions causing gelatin to become more soluble in water, hence the
> > > addition of chrome alum etc etc, and I suspect this is the root of
> > > the problem. Maybe one batch of subbing solution was not made
> > > correctly, or the a batch of staining solution is slightly more
> > > alkaline than usual.
> > >
> > > Hope this gives you a starting point.
> > >
> > > Cathy
> > >
> > >
> > > At 2:37 PM -0400 10/4/02, Knight, Brandon wrote:
> > > >I've been having a problem with 40um mouse brain sections coming off
of
> > > >subbed slides when placed in any liquid (for instance, when going
through
> > > >the staining process).  The edges may fold over, they may wrinkle, or
> > slide
> > > >off completely.  They were sufficiently dried (some for a couple of
> > weeks).
> > > >The slides that I'm using were subbed in August of 2000.  Is it
possible
> > > >that the gelatin has degraded and that these slides are no longer
subbed,
> > or
> > > >could there be another explanation?
> > > >
> > > >Brandon Knight
> > > >Children's Research Institute
> > > >Center for Genetic Medicine
> > > >Children's National Medical Center
> > > >Washington, D.C.
> > >
> > > --
> > > ---------------------------------------------------
> > > Cathy Gorrie
> > > Scientific Officer
> > > Neural Injury Research Unit,
> > > School of Medical Sciences,
> > > University of New South Wales
> > > Sydney, N.S.W. 2052
> > >
> > > Phone: 61-2-9385 2462
> > > Fax   : 61-2-9313 6252
> > > e-mail: c.gorrie@unsw.edu.au
> > >
> > >
>






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