About These Instructional Guides
The instructional guides listed below describe the basic operation of the Cytek Aurora spectral analyzers.
To complete the training, you should watch all the available videos—from System Startup to Shutdown.
For those who prefer a printable document, we included below a training guide in pdf format.
However, we strongly advise you watch the videos, even if you plan to rely on the pdf guide during your first experiments on Aurora.
To begin using the Cytek Aurora analyzers in the Flow Core at CHOP, please follow the guidance below:
- After watching the videos, you must create a 30-minute Full-Service reservation in iLab, preferably immediately before your first experiment on the Cytek Aurora.
During this 30-minute session, we will create your user account in SpectroFlo, answer any questions you may have, and help you begin familiarizing yourself with the instrument.
You may schedule your first experiment on Aurora either in unassisted or full-service mode.
- If you schedule a reservation on an Aurora analyzer in full-service mode, a Flow Core staff member will be available to assist you with your experiment. Full-service assistance is typically available Monday–Friday during business hours.
- For reservations scheduled in unassisted mode during business hours, we are typically available to answer specific questions, but our staff will not be available to provide extensive help with running your samples or troubleshooting your experiment.
- If you would like to practice on Aurora without using experimental samples, the Flow Core can provide a small set of reference control beads. Email flowcytometry@chop.edu
to request bead controls.
- We strongly recommend that your first experiment on Aurora include only reference controls and one or two samples. During this first experiment, you should focus primarily on familiarizing yourself with the device and the software, not on generating data.
- If you have any questions, please contact us at flowcytometry@chop.edu.
- 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.
- 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.
- 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.
- Export PDF files individually.
- Use Batch Analysis to export multiple PDFs.
- We recommend saving Experiment (zip) files.
- 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.
- After finishing samples, click Cytometer → Clean Flow Cell.
- Select Tube rack and follow the wizard.
- If last user, click Cytometer → Fluidics Shutdown.
- Select Tube rack and follow the wizard.
- 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.
Notes:
- The information in the pdf file is structured differently than in the video guides, but the general guidance is similar.
- All training materials above were prepared by the members of the Flow Core Lab at CHOP.
