Philippe Bourin writes: > >I have a Coulter ELITE ESP with 5 PMTs, a 488 nm Argon laser, a 633 HeNe >Laser. I whish to buy a 532 nm 200 mW solid Laser (Spectra physics). >Does any body use such a Laser with the two others? and what types of >problems should I have if I use FITC, PE Rhodamine and APC labelled >antibodies simultaneously? > Using more than 50 mW (or possibly even more than 25 mW) at 532 nm in an Elite will be counterproductive as far as PE excitation goes because even that much laser power will saturate PE. Also, rhodamine- and PE-labeled antibodies would probably be difficult to separate with optical filters. Also, 532 nm from the laser will crosstalk into the FITC channel. Just using the 532 and 633 nm lasers, it would be possible to use Alexa 532, PE, PE-Texas Red, PE-Cy5 and/or PE-Cy7 with the green beam and APC and APC-Cy7 with the red. -Howard
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