Re: Trouble mounting a hollow fiber membrane...

Posted by Martin Wessendorf-2 on
URL: http://confocal-microscopy-list.275.s1.nabble.com/Trouble-mounting-a-hollow-fiber-membrane-tp6188669p6189926.html

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Dear Roni--

On 3/19/2011 8:11 PM, Roni Prater wrote:

> I'm working with some hollow fiber membranes that are used in blood
> oxygenators. I want to quantify the amount of protein that has accumulated
> on these membranes after blood has passed through the system and to do this
> I need to image the membrane end on.
>
> The problem is that the fiber images out of the focal plane and thus I want
> to use a confocal for a 3D perspective. The confocal microscope is an
> upright scope and thus I need to "glue" my membrane, end on, onto a cover
> slip but I cannot find a good mounting substance that will harden fast
> enough to hold the membrane upright.
>
> I've been using ProLong Gold but it takes over night to harden and thus I've
> had to prop the membrane up using forceps overnight and it isn't very efficient.
>
> Are there any mounting media that could preserve fluorescence but hardens
> quickly? Does anyone have any ideas on how to mount this membrane for the
> confocal? Right now I'm using some rubber gaskets layered up with a hole in
> the middle to support the membrane ( it is a couple mm long). I super glued
> a coverslip to the top of the gasket and added the ProLong Gold to the well.
> The membrane is held down into the well overnight. The apparatus is then
> inverted and thus the coverslip is upright with the membrane "glued" to it.
> This seemed to work OK but I wonder if there is an easier way??

Could you embed the membranes in DPX (or other xylene-based mounting
medium)?  (You'd still have to hold the membranes up but for minutes,
not hours.)  Or alternatively, embed in agar, cut sections with a
vibratome and mount?

Good luck!

Martin



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