[Histonet] Sections floating off of slides

[Mary North]

Another step to help adhesion of the section is rehydration of the block
face before sectioning.  If the section appears to be chalky-white on
the slide it probably will not adhere well.  Take care not to
overrehydrate, especially brain, as it will swell and get mushy. If you
have a warming plate, it also helps after draining excess water from
under the section, to let it dry flat at 42 degrees C a few hours or
overnight.  The sections become translucent if they are properly dried.

Mary North, HT, HTL(ASCP)
Neuromuscular Laboratory
OHSU
Portland, OR

>>> Gayle Callis <gcallis <@t> montana.edu> 01/10/05 1:46 PM >>>

Several things could be happening

1.  No adhesive in either the waterbath - some add a few granules of 
gelatin as waterbath heats up or not using Plus Charge (silane or poly
l 
lysine coated) slides to keep sections adhered to the slide surface. 
If 
you use Plus charge slides, the water should be distilled and not tap
water 
as minerals in tap water in certain parts of the country will attract
to 
the Plus charge and negate section adherance.

2.  If you use ammonia water to blue the hematoxylin, it may be chewing
the 
protein sections off the slides aka alkaline protein hydrolysis which
makes 
sections release.  Change to Scotts tap water substitute to blue you 
sections.   This problem can occure even after oven drying of sections.
 If 
you use Ammonia water, it should be very dilute, just enough to blue
the 
section, change it daily since it is a cheap solution.  Sections can 
release with ammonia water bluing even IF you use slide adhesives.

3. Hopefully, your brain sections are very flat to perfect contact with
the 
slide. Sharp blades are a must.

At 01:31 PM 1/10/2005, you wrote:
>Hello All,
>I have encountered a problem H&E staining my paraffin embedded 8um
mouse brain
>sections.  The sections are floating right off some of the slides but
some of
>the samples are normal.
>The sectioning was done between mid-November 2004 through last week. 
The ones
>sectioned more recently were embedded simultaneously and seem to be
fine.
>I tried baking the slides at 55 C for an hour, the problem persisted,
but at a
>slower rate.
>Any tips to correct this problem would be greatly appreciated.
>
>Becky Proctor
>UNC-Chapel Hill
>
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Gayle Callis
MT,HT,HTL(ASCP)
Research Histopathology Supervisor
Veterinary Molecular Biology
Montana State University - Bozeman
PO Box 173610
Bozeman MT 59717-3610
406 994-6367 (lab with voice mail)
406 994-4303 (FAX)



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