CyFlow
™
Space | Operating Manual | March 2021
19
ii) No Errors Due to Calibration
Instability of beads suspensions over time or counting statistics of beads cannot influence
the counting results, since no beads are used.
iii) Less Preparation or Setup Time
The sample concentration is directly analysed by the FCM without any instrument
calibration or additional sample preparation step.
iv) Less Analysis Time
Concentration results for subpopulations, as defined by gates, are immediately displayed
on the instrument screen. No additional analysis steps, e.g. setting gates for beads, are
required.
v) Less Expenses
No reference beads required.
6.5 Performing True Volumetric Absolute Counting
Sedimentation and Count Time
Cells or other particles may tend to sediment inside the sample tube depending on their
density in relation to the surrounding suspension solution. Significant sedimentation can
take place within minutes. The CyFlow™ Space analyses the concentration at the location
of the sample uptake, where it can vary over time due to sedimentation or de-sedimentation
effects. Consequently, avoid too long count phases due to low sample speed
(e.g. below 2 µl/s). Count Time typically should not exceed 2-4 minutes.
1. Shake sample for re-suspension of particles, but avoid producing air bubbles on the
surface of the liquid.
2. Fill 850 µl of sample into a test tube (instrument setup should be set to: pre-run 2
sec, stabilize 2 sec).
3. Insert sample tube onto sample port.
The acquisition starts and performs:
Pre-run
stabilize
run
count: particles are counted during the count phase.
The acquisition stops automatically. Biosafety cleaning is done and Ready is
displayed when the system is ready to analyse the next sample.
4. Define subpopulations by regions and gates to analyse concentrations
(concentration will be displayed for each subpopulation).
During the count phase, user interference has to be avoided!
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