Performing True Volumetric Absolute Counting - Partec CyFlow space Instrument Operating Manual

Operating Basics
Precision and Reproducibility
Precision and reproducibility of
instruments can be demonstrated and checked
with Partec CountCheck beads (Product no. 05-
4010).
Benefits of True Volumetric Absolute Counting
Partec instruments can perform traditional Dual-
and Single-Platform counting and unique True
Volumetric Absolute Counting, the latter offering
unique benefits:
i) Precision
A high precision of better than 5% is guaranteed
by precise counting and mechanical volume
measurement. The counting reproducibility is
better than 2% rel. standard deviation.

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.
Depending as well on the particle size, significant
sedimentation can take place in terms of tens of
seconds or many minutes. The CyFlow
analyses the concentration at the location of the
sample uptake, where it can vary over time due to
sedimentation or
Consequently, avoid too long count phase due to
too low sample speeds. Count Time typically
should not exceed 2-4 minutes.
During the count phase, user interference must be
avoided!
Rev. 007 Date 2010-01-13
the Partec
®
space
desedimentation effects.
ii) No Errors Due to Calibration
Unstability 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.
1. Fill 830 µl of ready prepared sample into a
sample tube (instrument setup should be set
to: prerun 2 sec, stabilize 2 sec).
2. Shake sample for resuspension of particles,
but avoid air bubbles.
3. Insert sample tube onto sample port.
- the acquisition starts and performs
prerun
stabilize
run
count.
- particles are counted during the count phase.
- the acquisition stops automatically and ready
is displayed.
4. Define subpopulations by regions and gates
to analyse concentrations
- the concentration is displayed for each
subpopulation.
®
CyFlow space Instrument Operating Manual
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