> In my experience, the 3-color filter/dichroic system works just fine if you
 have
> reasonable staining to start out with, and if you're using the appropriate
> fluorophores.  The latter (for 3-color immunocytochemistry) would be
> fluorescein, lissamine rhodamine, and cyanine 5.18.  Secondaries conjugated
 with
> these reagents can be obtained from Jackson ImmunoResearch--they have them
> conjugated to donkey anti-rabbit, donkey anti-mouse, donkey anti-sheep, donkey
> anti-rat etc. etc. etc.  A good company to do business with, in my
> experience--Bill Stegeman is a prince.
> Martin W. Wessendorf, Ph.D.                                    (612) 624 9115
> Dept. Cell Biology and Neuroanatomy                       FAX: (612) 624 8118
> University of Minnesota                           [log in to unmask]
> Minneapolis, MN  55455  USA        [log in to unmask]
 
Thanks for this response.  I asked because we are getting gorgeous images
with Cy5 alone and with FITC or Bodipy and Rhodamine or Cy3 together, but
getting Cy5 in register with the others w/o such a cube is near impossible.
Thanks again.
 -mc