Hi Jacqueline, I suppose it is down to which lasers you have available. On a standard BD vantage the UV and 488 lines are most accessible so the obvious pairing is GFP with YFP or DsRed (RFP). If you have a tunable argon laser (as on a MoFlo) then the 457 line is fine to excite CFP and YFP pairs, although this will need compensating and fine tuning which filters you use. The BFP doesn't seem to be the prefered choice but it does allow for the largest spectral separation - BFP with GFP or YFP. Also don't rely too much on the "published" Excitation/Emission profiles of the GFP family, as they are most often different and require alittle bit of flexibility. Andy
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