Cytek Aurora
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Overview
Model: Aurora
Manufacturer: Cytek
Configuration
The CAT Facility has two identical Cytek Auroras in the main facility, which have been named Aurora Red and Aurora Blue to distinguish them. The PME Aurora is also managed by the CAT Facility, but located in the PME space. All Auroras have a number of exciting features:
- Spectral flow cytometer
- 5 lasers, 64 fluorophore detectors, 3 FSC/SSC detectors
- Detect a wide variety of fluorescent antibodies/dyes/proteins
- High sensitivity
- Cope with autofluorescence with the autofluorescence extraction feature
- Load samples from standard FACS tubes or a variety of plates
- Volumetric counter allows for cell count
Training Requirements
In order to use any cytometer in the CAT Facility without staff assistance, all users must complete the basic training outlined in the New User Checklist on the New Users page. The training for the Aurora specifically is:
- Flow Basics course (schedule online)
- Aurora training course
- Aurora hands on instrument training
- (hands on training for any other benchtop analyzers can be done at any time before you use another cytometer)
The Aurora training course covers all of the basic information needed to plan a successful Aurora experiment – creating proper controls, panel design tools, spectral unmixing, etc. Please contact Laura Johnston to schedule Aurora training.
Sample Preparation Tips
Accepted Tubes
The Auroras accept standard 5mL round bottom tubes (FACS tubes) as well as a variety of 96-well plates, including deep-well. It is possible to run 5mL FACS tubes with small volume tube inserts (AKA bullet tubes), however an extra setup step must be performed on the instrument to prevent damaging the sample introduction port (SIP).
Sample Volume
- Running samples on the Aurora is comparable to the Fortessas
- A minimum of 100 µL is reccomended, 150-300 µL is typical but use what is appropriate for the sample
- It’s best to bring concentrated samples, once you are at the cytometer you can easily dilute your tubes if they are running to quickly
- In general, run samples at less than 35,000 events/sec
Flow Rate
The volumetric counter allows for live monitoring of the sample flow rate. The estimated rates are:
- low: 15 µL/min
- medium: 30 µL/min
- high: 60 µL/min
Actual samples my vary. For example, many cell types run on low typically have rates around 9-11 µL/min.
Sample Loader
The Aurora-Red is equipped with an ASL Plate loader, while the PME Aurora has an AMS Plate Loader. See below for the features on both systems. The Aurora-Blue does not have a plate loader at this time.
Dead Volume
Dead Volume is the volume of the sample remaining inside the sample tube or plate well that cannot be aspirated into sample injection probe after each sample acquisition.
Resources
There are many resources in available in the Resources Library (located under the Resources menu). These can also be accessed in the module below.
Troubleshooting
How do I move the plate loader out of the way?
The Aurora failed QC
I have a low or decreasing event rate
There are no events displayed
Shutdown and maintenance procedure
Data won't transfer to the server
- The data should upload automatically every 30 minutes, so be sure that you gave enough time for the transfer to occur.
- Data should b exported to D:/CytekbioExport/FcsFiles/Experiments/FlowUser. Make sure that you did not export your files in another location – either somewhere else on the disk, or a sub-folder of FlowUser.
- In some case communication gets lost, contact the staff and we will alert our IT manager.
How do I export the Experiment file
From My Experiments in the SpectroFlo software, select your experiment and click Export. Navigate to D:/Experiment Server Transfer. The experiment file will be pushed to the server within 30min.
Contact the staff
Email cathelp@bsd.uchicago.edu to reach the CAT Facility staff.