Dear Flow Folks: I am a chemistry grad student, and my project is to sort genetically engineered libraries of E. coli. I am currently investigating ways to increase the rate of sorting on a commercial droplet sorter. Most of the commercial machines have a 76um orifice, which is much bigger than coli or yeast. I have noticed a reference from Shapiro's Practical Flow Cytometry (3rd) by Fellner-Feldegg on using a small 25um orifice. Is it realistic to fit a BD Vantage or a Coulter Altra with say a 20um orifice and a 100kHz droplet generation frequency, and thereby increase the effective sort rate? I am guessing that a cycle time of 5.5 microseconds means a maximum of 180,000 decisions/sec. This seems acceptable? I noticed that the Cytomation MoFlow uses droplet deflection technology, but would the BD or Coulter drop deflection plates result in acceptable purity and yield? Is jet-in-air required for these kinds of sort rates? Are there other considerations that need to be taken into account? I apologize for the ignorance, and thanks in advance. Mark Olsen BI/GG Group University of Texas at Austin
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