Re: Sections lifting off

From:Aidan Schurr

Poly-l-lysine slides should in theory stick down just about any kind of well processed tissue... so if tissues aren't sticking well, the first thing to look at is "are they properly processed?".  If so, lets look a little further afield.  How are you draining and drying the slides?  Any coated slides need to be *completely* drained before going near heat, otherwise small amounts of water get stuck under the section.  For ours we drain the slides vertically at room temperature for a minimum of 1hr, then into 37C incubator overnight.  Cook on at 60C for 20 minutes before staining the next morning (with the positive controls to ensure uniformity) to make sure they are stuck.  Still no luck?  Look at the water in your floating out bath... Are you using distilled?  Tap water just won't do in my experience.  Are you using any additives in the water bath?  These are an absolute no-no when it come to coated slides.  Eliminating the additive from our waterbaths about 12 months ago cured our lifting problems overnight.

Hope all this is of some help.  I know first hand just how frustrating it can be!

Cheers,
Aidan


__

aidan schurr  b.m.l.sc
section head, histology
hutt valley district health board
lower hutt
new zealand

aidan.schurr@hvh.co.nz
++64 4 570 9173 (direct)
++64 4 570 9214 (fax)

>>> LINDA MARGRAF MD  28/06/2002 >>>
Dear Histonetters
Here is a message Margaret is having trouble posting. She is on the
list so you may reply to the list or her directly at Mhorne@upei.ca.
Thanks.

My question is this.  How can I keep breast and eyeballs sections 
from lifting off the slides after heating them in the antigen retrieval

solution at 95 degrees for 20 min?  I have tried silanized and poly-l-
lysine slides with sporadic success. I am now about to try gelatin 
dipped .  Any ideas?   Thanks.
Margaret Horne 
Dept. of A&P,
Atlantic Veterinary College, U.P.E.I.
550 University Ave., Charlottetown,
P.E.I.,  C1A 4P3 
Canada





 

<< Previous Message | Next Message >>