Posted by
Armstrong, Brian on
Nov 01, 2012; 10:57pm
URL: http://confocal-microscopy-list.275.s1.nabble.com/TIRF-depth-calibration-tp5769934p7579267.html
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I am not sure how much wiggle you want, but PuraMatrix and Matrigel will both provide room for large colonies to form. You can obtain good 3D structure information using these in your set-up.
Cheers,
Brian Armstrong PhD
Assistant Research Professor
Director, Light Microscopy Core Facility
Beckman Research Institute
City of Hope
626-256-4673 x62872
-----Original Message-----
From: Confocal Microscopy List [mailto:
[hidden email]] On Behalf Of Glen MacDonald
Sent: Thursday, November 01, 2012 2:52 PM
To:
[hidden email]
Subject: Re: TIRF depth calibration
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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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> To join, leave or search the confocal microscopy listserv, go to:
>
http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy> *****
>
> 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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