Re: TIRF depth calibration
Posted by
Glen MacDonald-2 on
URL: http://confocal-microscopy-list.275.s1.nabble.com/TIRF-depth-calibration-tp5769934p7579265.html
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Hi,
Have you tried ~1.5% low melting point agarose? Maybe even lower concentration for your purposes. Use a coverslip bridge to create a spacer - use cyanoacrylate glue to stack 22 mm square coverslips to a desired depth onto a 50 or 60 mm coverslip. the top coverslip usually stays in place by capillary action.
Regards,
Glen
Glen MacDonald
Cellular Morphology Core
Center for Human Development and Disability
Box 357920
University of Washington
Seattle, WA 98195-7920 USA
(206) 616-4156
[hidden email]
On Nov 1, 2012, at 2:28 PM, Michal Opas wrote:
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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>*