Search the CONFOCAL archive at
http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal

Vectashield is a horrible mounting media for fluorescence because it is not
water soluble - so you wind up with water droplets throughout the media that
scatter light everywhere.  This might have added enough background to drown
out the real signal.  I would believe what I saw with the Prolong.  Better
yet, make your own mounting media:

20mM Tris pH 8.0 (final concentration)
0.5% N-propyl gallate
90% Glycerol
Warm to 37C and vortex to get into solution
Store at 4oC

Best, Jennifer

On 5/24/06, Xuhang Liu <[log in to unmask]> wrote:
>
> Search the CONFOCAL archive at
> http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal Hi everyone,
>
> I used vectorshield hard-set before but changed to prolong gold recently
> b/c the signal of my samples mounted in vectorshield often faded away in 2-3
> weeks. However, I was surprised to find that samples mounted in prolong gold
> gave me a different pattern from that using vectorshield hard-set. Before,
> my portein seemed to be diffusable in both the nucleus and the cytoplasm.
> Now, w/ prolong gold, my protein is still in both the nucleus and the
> cytoplasm, but it showed a net-like pattern in the cytoplasm, as if it was
> associated w/ some kind of cytoskeleton. I called the molecular probes, but
> they also haven't heard anything like this before. Does anybody have any
> idea what is happening or have met similar situation before? Thank you very
> much!
>
>
> -xuhang
>



-- 
Jennifer Waters, Ph.D.
Nikon Imaging Center at Harvard Medical School