Re: [Histonet] reversible tissue adhesion to slides

From:John Kiernan

How about this? (No, I've not tried it!)

1. Put a very thin tiny spot of cyanoacrylate (crazy glue)
on the slide, followed by the ganglion. Expose to water
vapour for a few minutes (to prevent drying, and to ensure
that all the cyanoacrylate polymerizes - the reaction is
catalyzed by water, metal ions etc).
2. Slice off the cells you don't want.
3. Shave the ganglion off the slide by sliding an
inclined razor blade along the glass.
4. Put the removed ganglion in culture medium or
whatever comes next.

I'd guess that the neurons contacted by cyanoacrylate
monomer would die. They might remain stuck to the
slide. All the other neuronal cell bodies and the 
central core of neuropil might be OK. Cyanoacrylate
glue is used for sticking unfixed specimens to the
chuck of a vibrating microtome, and it does hold
the specimens very strongly while they are being
sectioned immersed in liquid (water, saline etc
according to requirements). Cyanoacrylate adhesives
are used as tissue adhesives (instead of stitches);
they work well in wet places like the mouth, where
sticky tape would be no good. If the polymerized
glue is harmless in vivo it might also be harmless
if some bits of it end up in your culture.

Finally, if you can slice off the surface of the 
ganglion that you don't want, why can't you slice off and
collect the part that you do want to keep? That way,
no glue would enter the culture.

John Kiernan
Anatomy Dept, U.W.O.,
London, Canada.

"Janis C. Weeks" wrote:
> Dear colleagues,
> We are looking for a method to very firmly attach unfixed larval
> Drosophila ventral ganglia to a glass slide so we can slice off one
> surface of the ganglion (to remove neurons that we don't want; it's a
> long story; e-mail me if you want to know), to be followed by
> *releasing the attached tissue* from the slide and preparing primary
> neuronal cultures.  The tissue needs to be kept alive and happy
> under saline.  Polylysine doesn't hold tight enough for the slicing
> step (which we do with a fine blade).  Possibilities that have come
> to mind are using more adhesive slides (e.g., coated with
> aminosilane?), using some sort of double-stick tape, or a
> releasable glue (e.g., something like vetbond but that can be made
> to release?).  Obviously part of the difficulty is to release the tissue
> without destroying it.   Does anyone have a suggestion?  thanks!
> Janis Weeks
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> Janis C. Weeks, Professor of Biology
> Institute of Neuroscience
> 1254 University of Oregon
> Eugene, OR  97403-1254
> voice (541)346-4517, FAX (541)346-4548
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