Hi, Thanks to everyone who responded to our question about whether 100 mW of 488 from an air-cooled laser would be sufficient to do routine fluorescence on a Vantage. A resounding, yes, was the answer. Also, I mentioned (to someone) about the instability (I have heard about) of 5 watt argon lasers at under approx. 200 mW. The following technique (from one of the responders) is something that never occurred to me. Thought it might be of interest to other Vantage labs. "I've worked with three different Vantages at different core labs and I had no problem using only 15mW of 488 to excite cells. You may need to remove an ND filter and raise the gain setting in linear for FSC. Some lasers like a minimum amount of output to be stable. In the lab where this applied, I put a thick ND filter in front of the laser beam output to attenuate the beam intensity. The filter factor was around 3.14, so I ran the Enterprise I laser at 52mW to get effectively 15mW output. At power levels under 50mW, you could run PerCP without photo-bleaching." Ray Hester Univ. of South Alabama rhester@jaguar1.usouthal.edu
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