> I was wondering what you folks thought produed the best double label
> florescence for colocalization studies.  I tried TRITC and FITC and
> thought we had a little bleeding through.  Any other suggestions ???
 
The combination FITC - Texas Red is OK. You probably
will have to use an other emission filter for the red channel.
 
We are building a set-up for the simultaneous detection of three
different colours (FITC-TexasRed-Cy5) almost without any crosstalk.
This method is based on the modulation of the excitation beams and
demodulation of the detected signals.
 
 
Read about fluorophore choice:
Brelje, Wessendorf, Sorenson (1993) Multicolour laser scanning confocal
immunofluorescence microscopy: Practical applications and limitations.
Methods in cell biology vol. 38 p97-181
 
Read about intensity modulated beam scanning (IMS):
Carlsson, Aslund, Mossberg, Philip (1994) Simultaneous confocal recording
of multiple fluorescent labels with improved channel separation.
J. Microscopy 176: 286-299
Manders, Patwardhan, Carlsson (1995) Multi-channel confocal microscopy
by means of intensity-modulated multiple-beam scanning (IMS).
Zoological studies 34 supp I, p189
 
Erik Manders
 
____________________________________________________________________________
 
            Erik Manders                |
             Physics IV                 |       Phone:  +46-8-7906216
    Royal Institute of Technology       |       Fax:    +46-8-205609
         S-100 44 Stockholm             |       E-mail: [log in to unmask]
              Sweden                    |       Web: http://www.fysik4.kth.se
 
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