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Flow Cytometry

URMC / Research / Flow Cytometry / Services / Instruments / Analytical / Full Spectrum

Full Spectrum - Constantine, Sweetums and Link

The Aurora Full Spectrum Flow Cytometry instruments are new high end machines from Cytek Biosciences. The URMC FCR is fortunate to have three of these instruments in our facility.  The three options are Constantine (5 laser analyzer), Sweetums (5 laser analyzer) and Link Hogthrob (5 laser cell sorter).  The full spectrum cytometers use a new type of optical system capturing nearly the entire visible light spectrum and a process called spectral unmixing to resolve individual fluorochromes. The result is a system capable of performing 30+ colors. In addition to being able to resolve a large number of parameters this technology is also often able to distinguish highly overlapping options such as GFP and YFP or similar but not identical fluorochromes like Brilliant blue 515 and Alexa 488. This opens a new world of panel design options using combinations of fluors that we previously couldn’t reliably resolve. Using this technology we can also perform autofluorescence extraction allowing us to resolve negative or dim positive populations that previously were swallowed up by high autofluorescence in our cells of interest.

The two analytical systems also include integrated 96 well plate loaders allowing automated acquisition of a large number of samples.  The plate loader includes automated washing to reduce carryover and highly customizable sample acquisition to optimize for each experiment.

Constantine now has a small particle enabled SSC detector and is capable of measuring polystyrene beads as small as 100nm in size, which is a nice upgraded capability for those researchers studying EVs. Email us at FCRScheduling@urmc.rochester.edu for more information.

The cell sorter (Link) has the same 5 laser configuration as Sweetums and Constantine which allows direct transfer of whole experiments from one machine to the other.  An experiment that is tested and developed on these analyzers can then be transferred to the sorter and expected to work very much the same on all three instruments, saving time and reducing troubleshooting needs. 

Spectroflo Software

FCR offers the use of 2 Spectroflo software license keys to our Cytek Aurora investigators.  This will allow investigators to download and utilize the Spectroflo software on their own computers for unmixing and experiment creation purposes.  The USB license keys can be “rented” by emailing FCRScheduling@urmc.rochester.edu to make a request and arrange pickup.   The program is Win 10/11 only and we have an installation document to share, but your lab group will be responsible for the installation of the program on your own computers.  Click here for details.  

We will also continue to offer the Spectroflo analysis computer up here in the FCR hallway, but for those investigators that would prefer to be able to use the software on their own computer we will have that option as well.

General Information Before Proceeding

  1. Panel design considerations for the full spectrum analyzer are different than for traditional fluorescence flow cytometry.  Panels designed for any of the other analyzers will generally work on the Aurora, but to expand panels a discussion of options is recommended. Please see the documents listed below and contact  Matt Cochran if you'd like to schedule a time.
  2. Please request a training visit in PPMS if you'd like to schedule a time.  Training consists of two mandatory visits in front of the instrument for most users.
  3. Additional full spectrum cytometry and instrument specific information can be found at the links below.