CytoFlex Flow Cytometer Application Notes

Conclusion The introduction of Photodiode detectors expands the accessible electromagnetic spectrum for use in flow cytometry. In contrast to the photomultiplier tube, Avalanche photodiodes exhibit high quantum efficiency in the 800 nm (infrared) range of the spectrum. The CytoFLEX S platform offers the first commercially available flow cytometer incorporating both Avalanche photodiodes and an Infrared laser. Compensation challenges are reduced as a benefit of using the channels in the infrared range. No spillover from infrared emitting fluorochromes was detected in traditional channels, minimal spillover was detected in adjacent infrared channels only. Only minimal spillover was detected when two IR fluorochromes were tested in the adjacent IR Channels. To further demonstrate the usefulness, these new channels were used in biological applications. The addition of the 808 nm laser to the CytoFLEX S series provides additional fluorescent channels not only for use of viability dyes but also bright markers with minimal spectral overlap into traditional channels. Alternatively, the Infrared channels could be used for dimmarkers. The untouched nature of this part of the spectrum, either from the absence of auto-fluorescence or from spillover from other fluorochromes will maximize the sensitivity of detection. References 1. Mahnke, Y. D., & Roederer, M. (2007). Optimizing a Multi-colour Immunophenotyping Assay. Clinics in Laboratory Medicine, 27 (3), 469–v. http://doi.org/10.1016/j.cll.2007.05.002

2. Andersson, Baechi, Hoechl and Richter (1998). Autofluorescence of living cells. Journal of Microscopy , 191: 1–7. doi:10.1046/j.1365-2818.1998.00347.x

3. William G. Lawrence, Gyula Varadi, Gerald. Entine, Edward Podniesinski, and Paul K. Wallace. (2008). A Comparison of Avalanche Photodiode and Photomultiplier Tube Detectors for Flow Cytometry. Proc. SPIE 6859, Imaging, Manipulation, and Analysis of Biomolecules, Cells, and Tissues VI , 68590M (29 February 2008); doi: 10.1117/12.758958

For research use only. Not for use in diagnostic procedures. © 2017 Beckman Coulter Life Sciences. All rights reserved. Beckman Coulter, the stylized logo, and the Beckman Coulter product and service marks mentioned herein are trademarks or registered trademarks of Beckman Coulter, Inc. in the United States and other countries. SPHERO is a trademark of Spherotech, Inc. For Beckman Coulter’s worldwide office locations and phone numbers, please visit “Contact Us” at beckman.com FLOW-2674WP05.17

17