Hi Alice, Before you set the sweet spot, you need to compare the actual pixel number of drop 1 (on the right) to the target pixel number (on the left.) If these numbers aren't close you need to change the target number to match the actual number. If you don't do this, the sweet spot has a hard time determining where you want that break and will jump back and forth between the two breakoff points. Pam Shaw Fluorescence Cytometry Core Facilitator Center for Environmental Health Sciences The University of Montana 32 Campus Dr, Skaggs Building 052 Missoula, MT 59812 Ph (406) 243-4974 email: <mailto:pamela.shaw@umontana.edu> pamela.shaw@umontana.edu www.umt.edu/cehs <http://www.umt.edu/cehs/FluorCytomCore.htm> /FluorCytomCore.htm -----Original Message----- From: Furgeson, Alice N [mailto:Alice.N.Furgeson@pfizer.com] Sent: Wednesday, December 06, 2006 10:17 AM To: cyto-inbox Subject: FACSAria unstable drop 1 and gap Hello, I was wondering if any Aria users have experienced unstable drop 1 and gap even with the Sweet Spot on. I use a 100um nozzle. I have de-bubbled the filters, cleaned the flow cell with contrad 70, sonicated the nozzle and changed the O-ring. I have also tried using a different 100um nozzle and a 70um nozzle. The gap still fluctuates between 2 and 20 and the drop 1 goes between 100 to 300. The numbers just bounce everywhere. Any suggestion for why this is happening and any solution? Thanks, AliceReceived on Mon Dec 11 12:38:00 2006
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