Full Spectrum - Constantine, Sweetums and Link
The upgrade for Constantine has been completed! Please note: This will require recreating experiments and re-acquiring single stained reference controls for existing panels/experiments in some cases. In the cases that this experiment/panel has been run on Sweetums or Link previously however, you may be able to directly transfer your files/experiments over to Constantine. Please do reach out if you have any questions. Thank you.
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.
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.
General Information before proceeding
- 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.
- 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.
- Additional full spectrum cytometry and instrument specific information can be found at the links below.