Aurora Analyzer training – CHOP Flow Cytometry Core

Before your first experiment

To begin using the Cytek Aurora analyzers in the Flow Core at CHOP:

After watching the videos, create a 30-minute Full-Service reservation in iLab, preferably immediately before your first experiment on the Cytek Aurora. During this session we will create your SpectroFlo user account, answer your questions, and help you get familiar with the instrument. You may schedule your first experiment in either unassisted or full-service mode.
In full-service mode, a Flow Core staff member is available to assist you with your experiment. Full-service assistance is typically available Monday–Friday during business hours.
In unassisted mode during business hours, we are typically available to answer specific questions, but our staff cannot provide extensive help with running your samples or troubleshooting your experiment.
To practice without experimental samples, ask us for a small set of reference control beads at flowcytometry@chop.edu.

We strongly recommend that your first experiment on Aurora include only reference controls and one or two samples. Focus on getting familiar with the instrument and the software, not on generating data.

Questions? Contact us at flowcytometry@chop.edu.

Video guides

Click a title to play its video.

System startup

Check the tanks: ensure sufficient sheath fluid; empty waste tank.
Turn on the PC and cytometer.
Start SpectroFlo and log in.
Install a FACS tube with ~2 ml DI water on the SIT.
Wait for the cytometer icon to turn green. Loader must be on.

Experiment setup & Unmixing

Click Acquisition → New.
Name the experiment and add fluorochromes.
Select the appropriate Carrier Type.
Add the Reference Group and complete the wizard.
Record reference controls.
Click Unmix and adjust the gates in the unmixing wizard.
Click Live Unmix.

Gating & Acquisition Setup

Save the Unmixed worksheet.
Click Edit → Add sample group(s).
Adjust acquisition settings.
Add plots and gates as needed.
Run a sample and fine-tune gates.
The threshold value usually is left unchanged.
Record all samples.

Exporting data

Export PDF files individually.
Use Batch Analysis to export multiple PDFs.
We recommend saving Experiment (zip) files.

Importing & Duplicating experiments

In the Acquisition tab, click My experiments → Import.
Select the experiment to import and click Open.
Select Use existing item for Default Worksheets.
Duplicate with reference group data is commonly used.

Clean flow cell

After finishing samples, click Cytometer → Clean Flow Cell.
Select Tube rack and follow the wizard.

Shutdown

If last user, click Cytometer → Fluidics Shutdown.
Select Tube rack and follow the wizard.

Running samples from plates

Check that the loader and the cytometer are connected.
Start an experiment and select the plate in the carrier type
Continue with setting up the reference controls.
Drag-and-drop the controls in the plate layout to match their actual location.
Remove the FACS tube from the SIT and set the side arm to PLATES mode
Record the reference controls and unmix
Record samples, set plots and gates.
Export data.

For a printable guide, download the Aurora Analyzer Training Guide.

Download the training guide (PDF)

Notes:

The PDF is structured differently from the video guides, but the general guidance is the same.

All training materials above were prepared by the members of the Flow Core Lab at CHOP.