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Kevin Braeckmans wrote:
> As for custom filter sets, AHF Analysentechnik did a good job for us
> (www.ahf.de). In my experience, major players like Chroma don't even bother
> to respond to custom filter requests - and I have been told of similar
> experiences. I am just saying because it might save you some time.
> 

I have often ordered custom filters from Chroma, and they were very good about 
designing and supplying them.

Concerning choice of laser -- I find that fluorescence spectra are broad enough, 
   that you can do well with even large changes in excitation.  The published 
spectra may not be exactly what you are seeing in your system -- often they were 
measured in some organic solvent, not in cytoplasm, etc.

If you have a AOTF to select the laser line, then it probably needs to be 
reprogrammed for the new lines. On our FV1000 this is not difficult, and I 
assume the same is true for the other manufacturers, but you should check.
You will need the correct dichroic, but while you are waiting for it to arrive, 
try the flat 20/80 splitter (reflect 20% and transmit 80%). It will often do 
quite well, because you usually have power to spare from the lasers.  The 20/80 
splitter is great to have when you need a combination of laser lines that the 
manufacturer did not consider.

--aryeh
-- 
Aryeh Weiss
School of Engineering
Bar Ilan University
Ramat Gan 52900 Israel

Ph:  972-3-5317638
FAX: 972-3-7384050