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MACSQuant® Instrument quick guide
Setting up an acquisition
Setting up an experiment
on the MACSQuant® Instrument
Define an experiment
using the single tube rack
The MACSQuantify™ Software provides the user a simple
interface to program various sample parameters for data
acquisition. Follow this quick guide to see how to easily set
up a basic experiment for acquisition in the single
tube format.
Setting up parameters of the experiment tab
Follow along in figure 1.
1.Rack: Make sure single tube rack is selected.
2.File: (Optional) Change the file name.
3.Project: Select a folder path for files.
4.Sample ID: Provide unique identifier for sample.
5.Description: Similar to sample ID, add more detail.
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2.
3.
4.
5.
6.
7.
8.
9.
10.
Figure 1: ‘Experiment’ tab
6.Flow rate: Select a flow rate.
a.Low = 25 µL/min (for high cell densities
(~10⁷ cells/mL) or sensitive measurements)
b. Medium = 50 µL/min
c.High = 100 µL/min (for very low cell densities
(~10⁶ cells/mL))
Note: When choosing a fluidic speed it is best to keep the event/second rate below 10,000 for most accurate measurements. 1000–5000 events/second
is ideal.
7.Mix sample: Select if sample should be mixed prior
to measurement.
8.Mode: Select inter-sample wash mode (fast, standard,
or extended). Standard is default by factory setting.
9.Uptake volume: Enter volume for measurement.
10.
Sample volume: Enter sample volume.
Note: If ‘mix sample’ is selected the sample volume
must be entered accurately.
Setting up custom settings of the experiment tab.
Follow along in figure 2.
1.Instrument settings: Select saved PMT and
compensation settings for sample(s).
Note: If current live instrument settings on the
instrument are valid for the sample, there is no need
to select a setting under this tab.
2.Analysis template: Select a saved plotting/gating
strategy for sample(s).
Note: Alternatively a new analysis template can be
created for this sample. Refer to section below or
to the user manual.
3.Gate: (Optional) Select a live gate or stop gate to define
population for analysis.
4.Events: (Optional) Define event limit for analysis gate or
sample. This is often used when a stop gate is active.
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2
Selecting a display layout for analysis
Note: This is NOT necessary if a pre-defined analysis
template is selected within settings tab.
1.Select a plot layout by clicking on the icon.
2.From the pop-up window, select a layout suitable
for sample analysis (figure 3).
Note: Each specific plot can be changed to display data
as a dot plot, density plot, histogram plot, or statistics
table. Click on the ‘i’ button to select different options.
3.Change axes to display the optical channels suitable
for sample analysis by clicking on the axis label and
choosing from the drop down list.
4.Use channels tab to adjust axis scales if necessary
to be suitable for sample analysis (e.g. FSC in linear,
SSC in linear, and fluorescence channels in HLog).
5.Gates can be drawn on the plots by clicking one of these
icons and clicking and dragging on the
appropriate plot.
6.Annotations can be modified to fit staining panel,
by opening annotations tab (figure 4) and typing in
appropriate fluorochromes or dyes.
Note: Annotations may not appear on plot display
windows until acquisition starts.
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Figure 2: Settings tab under ‘Experiment’
Figure 4: ‘Annotations’ tab under ‘Experiment’
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Figure 3: Selection of plot display layout