[Histonet] Re:sealing slides with diluted permount
gcallis <@t> montana.edu
Mon Dec 19 13:15:36 CST 2005
We buy cheap clear finger nail polish and dump it out!!! Rinse out the
little polish bottle and brush with acetone and let it dry. Put Permount
in the bottle, add some xylene or toluene (if toluene based). When you buy
fingernail polish, look at the type that is for base or top coat, tip it so
it flows and look at consistency or thickness of this polish. This is the
thickness you want, I don't bother with proportions, as you can use thick
goo if you want - thinner is easier to apply and it dries almost instantly.
To apply the diluted Permount with the little polish brush to edges of
coverslip, generally a put a little over the edge to see a good seal. If
you pipette it, you will have a mess going everywhere. You can also use
an artists brush like we use for sectioning, and wash it with solvent
afterwards. However the fingernail polish brush is ideal, stiff, short
bristles that you can control application of media which is now stored in
the little polish bottle.
Reusing the fingernail polish bottle is ideal. I generally do two edges
over the glass slide, and not bother with edges of coverglass on outside
edges of slide, if you have 24 x 24 coverglass, you have to be careful not
to get media on bottom of slide. Some people might like a 22 x 22
coverglass or 22 x 30, or 40 so you can seal ALL edges.
At 11:51 AM 12/19/2005, you wrote:
>I have some slides of GFP expressing tissue. I am using the prolong
>gold mounting media from Molecular Probes to mount the coverslips but
>now I have to seal them. I have read that the best way to do this is
>with diluted permount. Unfortunately I have never sealed slides
>before and am having a hard time visualizing the process.
>What is a good starting dilution for the permount? I was thinking
>1:10 with xylenes to start, but this is just a guess. How exactly do
>you apply the permount? Can you just pipet it around the edges or do
>you need to brush it on in some manner? Does it have to be flush
>with the coverslip or can you have some buildup?
>Any advice would be appreciated.
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Veterinary Molecular Biology
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