Here is a question for the group. We are currently using a Coulter Elite ESP sorter with an air cooled argon laser and a water cooled Inova 90-5. We are working with particles that have FITC and another similar particle that has Rhodamine attached to the particle. When we use our air cooled laser at 10 mWatts of 488 we see about a three fold increase in fluorescence of our rhodamine particle and about 2 log increase with the FITC. If I up the air cooled argon laser to 40mWatts we see a 100fold increase with the Rhodamine and a three log increase in fluorescence with the FITC. I assumed that the rhodamine, as well as the FITC, was brighter simply because they were now absorbing more 488nm light. Here is my first question. Is our laser putting out only 488nm light or are other wavelengths coming out as well (such as the 514 or 528 lines and are these wavelengths resposnsible for the rhodamnine fluoresing brighter)??? As I understand lasers, the light output is monochromatic and coherent. The second question: I was thinking I could increase the fluorescence of the Rhoadmaine by using a wavelenth of light closer to its excitation maxima(around 550). Therefore, i want to use the air cooled laser at 488nm and the water cooled innova 90-5 at 514 or 528. Has anyone used a 488nm and a 514 or 528 at the same time? Does it work well with the gated amp?? Could I get by with just using the 514 for both fitc and Rhodamine? I appreciate the input about the output:) Mike
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