FACScan side scatter

From: Immuno Flow (Immuno_Flow_at_APPLIC1@ccmail.llu.edu)
Date: Thu Dec 21 1995 - 14:16:02 EST


     I still believe this "blip" is air bubbles because we "supposedly" 
     have dedicated lines and surge protectors on all of our equipment.  
     But...who knows?
     
     We're currently using Lysis II on our FACSort, but, have used the 
     Consort 30 and Cytomation and Mac software on our FACScans and have 
     still seen this "blip" when acquiring.  To reduce air in the lines, I 
     try the typical restart, gate out, drain/fill (for complete shifts in 
     populations between tubes).  We also use Isoterge from Baxter 
     Scientific (normally used for coulter counters) which my previous boss 
     seemed to think it reduced surface tension therefore reducing air 
     bubbles in the system.  We continue to run 5 min. of each isoterge, 
     10% bleach, and distilled de-ionized water at the start and end of the 
     day.  If I have this complete shift in populations, mainly noticeable 
     in phenotyping, and drain/fill still doesn't purge the bubble, I run 
     isoterge then water for 10 min. each and that usually helps.  (Note: 
     Isoterge is a detergent which could create bubbles if you were to 
     drain/fill with out washing the lines with bleach and water.)
     
     There is still the possibility of the sample contributing to air 
     bubbles. Your best bet is to restart if you can't gate it out either 
     on acquisition or analysis.
     
     Judy Folz
     Immonology Center
     LLUMC


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