Which aqueous mounting media are you using and how do you coverslip?
How are you doing the coverslipping and which mounting media?
Try a more generous drop os aqueous mounting media after wiping away excess
water around section. I wonder if the water is degassing? AquaMount
(Shandon Aqueous mounting media) is our favorite. Some medias have a
thinner, watery consistency - these tend to give us more problems. DAKO
has GlycerGel that is heated, but when applied to a dried section, was
excellent, plus it produces a hard set, no messy goo coming out around
edges or unstable coverslip.
If we draw circles around section with a hydrophobic barrier pen, the flow
of media is impeded by hydrophobic goo - we get more bubbles or bubbles
form near/on goo. We prefer to draw a line above and below section to
retain reagents during IHC. It is easier to blot reagents from tilted slide
with a squared off area/line than from circle, and when coverslipping, the
edges on either side of section are open, clear of goo - media flows evenly
from one side of slide to other.
Research Histopathology Supervisor
Veterinary Molecular Biology - Marsh Lab
Montana State University - Bozeman
S. 19th and Lincoln St
Bozeman MT 59717-3610
406 994-6367 (lab with voice mail)
406 994-4303 (FAX)
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