Hi Rich, The 200um nozzle is fine to use, although strangely enough we have had more success with the 300um nozzle in the last year. The reason for this in my opinion is variability in the "quality" of the orifice of the nozzle, as you seem to be able to get a good one, which gives good reproducible break offs and others which are just plain painful to use. For the 200um try 4.5 PSI and just wind the frequency down until in gets in the "scare the dogs" range and find a usable breakoff point. Remember that bubbles are the killer for large nozzle sorting, so go to some lengths to make sure you have minimized them in the lines, de-gas the sheath if you want to do a better job of this, and make your own call on hooking the sheath tank up via the alternate port on the tank, although personally I think this makes the bubble issue worst rather than better. If you really want to get things right, put another pressure regulator in-line (with independent gauge) with the sheath pressure regulator to give you the fine control you need hit the same sheath pressure day after day. The standard regulator is not really designed with fine level of control this type of work requires. Last, but not least, be patient, it takes a while for the system to settle, and you're not going to sort vast numbers of cells. Hope this helps, Geoff -- Geoffrey Osborne Director of Flow Cytometry, The Queensland Brain Institute /Australian Institute for Bioengineering and Nanotechnology, The University of Queensland, St Lucia, QLD, 4072, Australia Ph (61) 07 33469912 email g.osborne@uq.edu.au On 26 Jun 2006 at 9:48, Konz, Richard wrote: > > > Greetings everyone: > Is anyone out there sorting on a Vantage SE DiVa with a 200 um nozzle? Any tips or words of wisdom > would be appreciated. > Best regards to everyone, > -Rich > >Received on Wed Jun 28 12:58:00 2006
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