Gene Pizzo, M.S Manager, Flow Cytometry Facility UCONN Health Farmington, Ct. 06030 860 679-7567 http://flowcytometry.uchc.edu <http://flowcytometry.uchc.edu/> ________________________________ From: Pizzo,Eugene Sent: Thu 12/6/2007 18:37 To: cyto-inbox Subject: RE: 6 side streams in ARIA?!? Hi Vinko, That's a really cool picture and in some sense it shows the future of flow cytometry as the engineers learn to add more streams AND more colors to sorting but what you have is not six streams but four streams with center stream fanning. It is the result of a drop drive frequency that is "inadequate" to clarify drop separation. On the ARIA large nozzle sizes are somewhat unstable and require a fine tuning of nozzle position, drop drive frequency, amplitude, and pressure. Use the built-in "low" setting only to ballpark. Then begin by adjusting pressure and drop drive frequency to be certain you're getting good separation of drops and merging satellites. Because the separation of your streams looks otherwise so good I'd guess it's just a minor frequency change to tighten your center stream. You're plate voltage - at 5000 volts - is too high for a large nozzle however, that could be "stretching" your streams. I only use such a high voltage for higher pressures/smaller nozzles. I run the 100um nozzle closer to 2,500 volts. Don't bother with the 2nd, 3rd, and 4th drop adjustment - strangely I've never seen this have any effect since the old Vantage/SE days - since we've gone digital if you need to use the drops adjustment it's a good bet you're in the wrong frequency. Keep it at 20,10,5. Gene Pizzo, M.S Manager, Flow Cytometry Facility UCONN Health Farmington, Ct. 06030 860 679-7567 http://flowcytometry.uchc.edu <http://flowcytometry.uchc.edu/> ________________________________ From: Vinko Tosevski [mailto:vinko.tosevski@medri.hr] Sent: Wed 12/5/2007 17:11 To: cyto-inbox Subject: 6 side streams in ARIA?!? Hi, My department recently bought an Aria and I was among the first ones to go through operator's training for it. So, although I have previous flow experience, I am largely a beginner in the sort world. Today I tried to sort some cells using 100u nozzle and LOW sort mode (for the first time; most of the time we use 70u nozzle and HIGH sort mode). The stream looked OK but, when I applied the voltage and used TEST SORT button, I got 6 side streams instead of 4!??! (I will try to attach a picture but I've never seen any attachment going through) Is there a simple explanation to this phenomenon and how can I resolve this? When I move the sliders used to adjust the position of side streams, the outer two side streams on each side respond. But, the center stream splits/gets 2 side streams of its own after applying a voltage, and they are inresposive to the movement of any sliders. These two are not too much apart but are clearly visible as separate streams in camera window... Any comment is more than welcome. Sincerely, Vinko ---- Vinko Tosevski, MMolBiol Research fellow/Teaching assistant, Department of physiology and immunology, Rijeka Medical School, Ul. b. Branchetta 20, HR-51000 Rijeka, Croatia Tel: +385 51 651 192 Fax: +385 51 675 699Received on Fri Dec 7 14:38:00 2007
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