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Setting Up An Acquisition: Best in Class

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83 views2 pages

Setting Up An Acquisition: Best in Class

Uploaded by

mhd
Copyright
© © All Rights Reserved
We take content rights seriously. If you suspect this is your content, claim it here.
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Best in class

MACSQuant® Instrument quick guide

Setting up an acquisition
Setting up an experiment Define an experiment
on the MACSQuant® Instrument using the single tube rack
The MACSQuantify™ Software provides the user a simple Setting up parameters of the experiment tab
interface to program various sample parameters for data Follow along in figure 1.
acquisition. Follow this quick guide to see how to easily set  1. Rack: Make sure single tube rack is selected.
up a basic experiment for acquisition in the single 2. File: (Optional) Change the file name.
tube format.
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.
6. Flow rate: Select a flow rate.
a. Low = 25 µL/min (for high cell densities
1. (~10⁷ cells/mL) or sensitive measurements)
2. b. Medium = 50 µL/min
c. High = 100 µL/min (for very low cell densities
3.
(~10⁶ cells/mL))
4.
 Note: When choosing a fluidic speed it is best to
5. keep the event/second rate below 10,000 for most
6. accurate measurements. 1000–5000 events/second
is ideal.
7. Mix sample: Select if sample should be mixed prior
7. to measurement.

8. 8. Mode: Select inter-sample wash mode (fast, standard,


or extended). Standard is default by factory setting.
9.
9. Uptake volume: Enter volume for measurement.
10.
10. 
  Sample volume: Enter sample volume.
Note: If ‘mix sample’ is selected the sample volume
Figure 1: ‘Experiment’ tab must be entered accurately.
Setting up custom settings of the experiment tab. Selecting a display layout for analysis
Follow along in figure 2. Note: This is NOT necessary if a pre-defined analysis
1. Instrument settings: Select saved PMT and template is selected within settings tab.
compensation settings for sample(s). 1. Select a plot layout by clicking on the icon.
Note: If current live instrument settings on the 2. From the pop-up window, select a layout suitable
instrument are valid for the sample, there is no need for sample analysis (figure 3).
to select a setting under this tab.
 Note: Each specific plot can be changed to display data
2. Analysis template: Select a saved plotting/gating as a dot plot, density plot, histogram plot, or statistics
strategy for sample(s). table. Click on the ‘i’ button to select different options.
Note: Alternatively a new analysis template can be
created for this sample. Refer to section below or 3. Change axes to display the optical channels suitable
to the user manual. for sample analysis by clicking on the axis label and
choosing from the drop down list.
3. Gate: (Optional) Select a live gate or stop gate to define
population for analysis. 4. Use channels tab to adjust axis scales if necessary
to be suitable for sample analysis (e.g. FSC in linear,
4. Events: (Optional) Define event limit for analysis gate or
SSC in linear, and fluorescence channels in HLog).
sample. This is often used when a stop gate is active.
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
1 appropriate fluorochromes or dyes.
 Note: Annotations may not appear on plot display
2 windows until acquisition starts.
3

Figure 2: Settings tab under ‘Experiment’

Figure 4: ‘Annotations’ tab under ‘Experiment’

Are you in need of additional assistance?


Visit www.miltenyibiotec.com/locations to find
your nearest Miltenyi Biotec contact.

Figure 3: Selection of plot display layout


130-106-082

Miltenyi Biotec provides products and services worldwide. Visit www.miltenyibiotec.com/local to find your nearest Miltenyi Biotec contact.

MACS, the MACS logo, MACSQuant, and MACSQuantify are registered trademarks or trademarks of Miltenyi Biotec GmbH. Unless otherwise specifically indicated,
Miltenyi Biotec products and services are for research use only and not for therapeutic or diagnostic use. Copyright © 2014 Miltenyi Biotec GmbH. All rights reserved.

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