[Histonet] re: triple immunofluorescence for apoptosis (blue fluorophore?)

[Britt Tracy]

 Good morning!

I'm looking for advice on an antibody conjugated with a blue fluorophore
that could be used to tag for apoptosis (on p75 receptors on cholinergic
neurons in the rat basal forebrain).

We use a Goat X ChAT primary (1:100) and a Rhodamine-conjugated Donkey X
Goat secondary (1:250) on the slices.

The slices already contain GFP expressed by the viral vector we infused into
the basal forebrain.

So, when we image the sections we already have a green fluorophore and a red
fluorophore. Is there a blue fluorophore tag for apoptosis that anyone has
used which would be biochemically compatible with the antibodies above?


Is there another fluorescent option? Internet-searching and
literature-reading has not gotten me anywhere.

Our scope is a Leica DM4000B equipped with a Leica DFC 425C camera.



Thanks for your time; any advice or insight would be much appreciated.


Sincerely,

Britt Tracy (undergraduate researcher at Temple University)