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Dingyi Xu,
All three of the sorting problems you mention probably have the same
root cause; fluidics instability due to air bubbles. In my experience
with the Elite, air bubbles get caught in the flow cell tip near the
sample insertion rod exit point. These bubbles are not visible since they
are below the opaque region of the flow cell tip assembly. The Clear
function by itself will frequently not eliminate these air bubbles and,
in fact, may introduce more bubbles. The way I purge the flow cell tip of
air bubbles is to immerse the flow cell tip up to the top of the silvered
concave lens in a 50 mL conical tube filled with Coulter Clenz. I do this
while the fluidics system is operating and the flow cell tip is fully
assembled. Once immersed, I put the fluidics system in Vacuum mode and
then in Clear mode. You will see the bubbles rise. Repeat this procedure
until no more air bubbles rise in the flow cell. Return the fluidics to
Standby mode and remove the Coulter Clenz. Gently dab the flow cell tip
with a cotton swab to dry, do not rub since you may scratch the surface.
In my experience, the sorting oscillator on the Elite takes a good two
hours to stabilize from a "cold" start. Our sorting parameters are 30 kHz
Frequency, 75% Drive, 100% Deflection. We gennerally sort 3 Drops for
highest purity. Drop delay has been stable for one year at 41.0. Phase
changes are rarely needed.
Good luck,
Mark Rehse
Rehsema@CellPro.CellPro.com
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