CyFlow
™
Space | Operating Manual | March 2021
25
NOTICE
Parameter names cannot be changed after an acquisition.
Parameter names
Check if the parameter names are according to your optical setup. Typical names are
shown in the example above. The parameter names are displayed on the histogram axis
together with a parameter label. To change parameter names, click into the fields and enter
the names by using the keyboard. Do only use letters and numbers and no special
characters like \-_./><+*() to name the parameters. We do recommend to maintain the pre-
set parameter selection (do not enable) and parameter names. Modify parameter labels in
instrument settings, only.
Pulse Property Selections
In the CyFlow™ Space Signal Peak height is used as default to determine signal size for
all detectors. Next to signal height, peak area or peak width can charaterize the signal peak.
It is possible to activate in addition to the Signal Peak height peak area and/or peak width
for up to 2 detectors. This can be used to discriminate in the DNA parameter between single
cells and aggregates, e.g. by a peak hight versus peak area dot plot.
Low Pass
The electrical low pass filter smoothes the signals from the detectors by averaging the
signal over a given time. This feature is only required for UV-LED derived signals.
2 Wavelengths Signal Delay
Light beams from different lasers are directed to different positions (spots) of the flow
cuvette, see chapter 3.2 Flow Cytometry Analysis. Therefore, all laser derived signals from
a laser at a spot other than spot no. 1 need to be analysed with a time delay of 50 µsec.
9.1.1 Trigger
Leading Trigger
In order to discriminate particles of interest e.g. cells from other particles e.g.
cell fragments or nutrition particles in cell culture, a proper trigger has to be
chosen. In the CyFlow™ Space, any parameter can be used as leading trigger.
The trigger is indicated by the black Asterix sign on green background.
Selecting one leading trigger parameter will automatically deactivate the
previously selected leading trigger. Only one leading trigger can be selected
at a time.
Frequently, the forward scatter parameter (FSC) or, preferentially for smaller
particles, the side scatter parameter (SSC) is used to trigger on all particles
above a certain size range. However, especially for very small particles, e.g.
microorganisms, triggering on a fluorescence parameter can be more efficient.
Selecting a parameter as leading
trigger parameter
means: Only particles that deliver a
sufficient signal above the lower level threshold on that parameter will be acquired. Other
non-triggering parameters are aquiring signals only for particles that generated a valid
trigger signal (above threshold). All other particles will not be “recognized” by the
instrument. The trigger parameter can be used for an efficient exclusion of unwanted
particles from the analysis.
Summary of Contents for CyFlow Space
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