Trouble mounting a hollow fiber membrane...

[Roni Prater]

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Hey all...

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??

[Martin Wessendorf-2]

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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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