[Histonet] confocal condition for Cy2 and Cy3 IHC on plants

From:Moran Oliva

Hello all!
I've just joined this forum. It is very useful. 
I've started a preliminary assay of Immunohistochemistry on Arabidopsis tissue (Steedman's protocol) using my GFP-protein transgenic plants as a positive control (to confirm the method is working for me).
The anti-GFP antibodies I used were either from mouse or from rabbit. So when I used the Cy2 goat anti-mouse I needed to use the same emission spectra as for the GFP protein itself-  Does anyone knows whether the GFP survive all this process? does anyone has any good advise of detecting the IHC signal and not theprotein GFP signal? the second channel I used was GFP autoflouresence...
Does anyone know what are the confocal setting details for Cy3 in plant tissue? I've tried to detect the GFP in IHC using rabbit GFP protein, so my second antibody is Cy3 conjugated ( I know the excitation and emission spectra, but the autoflourescence is very close to its wave lengths-do I need to use a second channel for autoflourescence in this case too?

Thank you very much
looking forward for your advices
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