Justin, It sounds like you would like to measure FSC-W. The following is a simple way to get FSC-W off the FACScan as long as at least one of the fluorecent dectectors is not being used. First you have to take the cover off the FACScan. Take the signal lead (not the high voltage lead) off the FSC photodiode. Next take the siganl lead off the Fl-1 or Fl-2 or Fl-3 (Fl-X) pmt and attach the signal lead from Fl-X to the FSC photodiode. Now your FSC signals are routed through the FL-X electronics and FLX-W is really FSC-W. Remember to threshold on Fl-X and that the FL-X gain (not voltage) is used to adjust the relative FSC signals. Good luck. Glenn MIT >Greetings, > >We are attempting to use time of flight (pulse width) on a FACScan to >accurately measure cell size on different cell lines. We are trying to >avoid the sticky business of using forward scatter with bead standards to >establish a curve. That method has its own set of problems. As FACScan >users are aware, pulse width is only available on the three fluorescent >channels. We initially tried using FDA to get a decent fluorescent signal >for the pulse processor to derive the width signal. Our results did not >correlate with known cell sizes. The trends were all wrong. Has anyone >else successfully used the time of flight on a FACScan for such >measurements? Thanks for your input. > > >------------------------------------------------ >| Justin Fishbaugh | >| University of Iowa | >| Flow Cytometry Facility | >| 48 EMRB | >| Iowa City, IA 52242 | >| | >| justin-fishbaugh@uiowa.edu | >| http://www.medicine.uiowa.edu/flowcytometry/ | >------------------------------------------------
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