Thermo Scientific gibco CTS Rotea User Manual page 83

Note: Speed represents a combination of flow rate and G force where the ratio of G force : Flow Rate is
roughly constant for the same fluidized bed conditions. For example, 1,000 G : 50 mL/min is equivalent
to 2,000 G : 100 mL/min.
The following section describes a method for determining suitable elutriation speeds for processing
PBMCs but the same method could equally apply to other applications.
Elutriation process development
CTS ™ Rotea ™ Counterflow Centrifugation System Process Design User Guide
Image
Chapter 4 Protocol Building Blocks
Details
Set up a 2-port input bag between lines G and D.
Insert a sampling port in line G.
Establish a bed of PBMC's using recirculation in the
planned media with Skip Button as the only trigger.
Manually adjust the pump flow rate or centrifuge
™
speed using the Rotea GUI, allowing a few seconds
for the system to reach a steady state condition.
Collect a sample of the elutriation output from the
sampling port in line G using a syringe. Repeat
at different settings to determine the first break
point i.e. when the smaller lymphocytes are being
elutriated. Continue to increase the flow rate or
reduce the G force until the larger monocyte cells
are detected, defining the second break point.
4
Elutriation
83