The Flow Cuvette - Partec CyFlow space Instrument Operating Manual

Appendix

The Flow Cuvette

Note
The flow cuvette is "the heart" of your flow
cytometer. It is responsible to guide cells and
particles through the center of the illumination
spots with a micrometer precision. The flow cell is
manufactured with highest possible precision and
should be treated like that. Any deformation will
degrade the flow
irrepairably unusable.
Waste outlet
Quarz cuvette
Sample nozzle screw
Sample inlet
Fig. 12: Flow cuvette with sample, sheath fluid
and waste connections.
Note
Air bubbles usually enter the flow cuvette through
the sheath fluid inlet. Bubbles might stick inside
the conical section above the sample nozzle and
do not enter the flow channel of the quartz cuvette
segment. The laminar flow of sheath fluid and
sample flow as well as hydrodynamic focusing are
disturbed and broad peaks appear in the
histograms.
Note
Do not readjust the laser or laser optics.
DANGER
Warning: Do not look into the laser beam. Use
eye protection glasses of filters or block the
laser beam.
Rev. 007 Date 2010-01-13
precision and make
Sheath fluid inlet
Cleaning of the flow cuvette
For cleaning the flow channel use Partec
Cleaning Solution (Product no. 04-4009) filled into
a standard sample tube and connect to the
sample port 2-3 times. Then perform additional
cleaning cycles with destilled water filled into a
fresh sample tube.
Do not use mechanical wires or tools for cleaning
or declogging the flow cell.
it
Removing Air Bubbles from Flow Cuvette
If you notice unsufficient peak resolution during
sample run remove your sample and activate
CLEAN several times. In this way the flow cuvette
is back flushed with sheath fluid. Restart the
sample and observe peak resolution again. If the
results are still insufficient, run Partec Calibration
Beads (e.g. 3 µm Calibration Beads). Compare
the result with the exemplary data files located in
the FloMax/QC
directory and observe the flow monitor (CCD
camera). A single bright spot should appear in the
centre of the visible field. If this is not the case air
bubbles or dirt inside the flow cell may cause the
disturbance. As first easy approach to release
bubbles during system run attach a tube with
Partec Cleaning Solution and pinch first the
sheath and then the waste tubing several times
to remove air bubbles through the flow channel
into the waste.
In case air bubbles are still persistent please
apply the following procedure:
1. Select the instrument settings:
Sample prerun = 15 sec
Stabilizing time = 5 sec
Automatic stop = 15 sec
2. Place an empty sample tube on sample
port
3. Press START and wait till automatic stop
4. Place a tube with 1.5 ml Partec Cleaning
Solution on the sample port and repeat
step 3.
5. Reset instrument settings to normal
values
If analysis with Partec Calibration Beads does not
result in acceptable signal distributions an
incubation
Decontamination Solution is required. Run the
system with 1.6 ml Partec Decontamination
Solution. During system RUN pinch the sheath
fluid tube for 5 seconds. Stop the instrument while
keeping the tube pinched by pressing STOP.
®
CyFlow
or FloMax/Calibration
of the flow cell
space Instrument Operating Manual
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with Partec
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