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Dear Michal,
One option is to put drops of nail polish on the coverslip, let them dry, then they act as spacers to prevent the coverslip crushing your cells.
Or any other solid inert things of about the right dimensions - we often use slices of coverslips as spacers, slice them with a diamond pencil. If they're not deep enough, we just pile 2 or 3 on top of each other.
cheers,
Rosemary
Dr Rosemary White
CSIRO Plant Industry
GPO Box 1600
Canberra, ACT 2601
Australia
T 61 2 6246 5475
F 61 2 6246 5334
________________________________________
From: Confocal Microscopy List [
[hidden email]] On Behalf Of Michal Opas [
[hidden email]]
Sent: Friday, 2 November 2012 8:28 a.m.
To:
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Subject: Re: TIRF depth calibration
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Dear Listers,
My search of archives leads me nowhere hence this question:
we need to mount spherical cell aggregates (mouse EBs) circa 300 µm in
diameter in a mountant that would let them wiggle while focused up and
down and, importantly, prevent coverslip from moving and shearing them
to shreds. Our commercial mountant (DAKO) does not cure. Nail polish
remedies this a bit but I'd hope for an improvement with a mountant that
would gel (solidify) in a decent time.
Thank you very much in advance!
Michal*
<
http://www.utoronto.ca/mocell>*
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