Spectral Flow Cytometry - the real story

NEWS FLASH: Spectral Flow Cytometry Article published April 30, 2019


Gerald Gregori's Spectral Cytometry page is here 

Before Spectral Flow Cytometry: For many years we have focused on polychromatic flow cytometry as being the fundamental tenet of flow cytometry. We moved from 1 or 2 colors, to three colors to 8, 12, 18, 27 and more. But the technology never really changed. All we did was add more PMTs and select more lasers. While this works pretty well, there are many complications that make doing high color polychromatic flow cytometry very difficult. There may well be a better way!


Spectral flow cytometry is taking off. We would like to set the record straight. The core development of spectral flow cytometry was first presented by the Purdue group at the 2004 ISAC meeting in Montpellier and published by our group in a biophotoncis International paper here in 2004. A paper was published in SPIE in 2005. An additional paper "Multispectral FLow Cytometry: Next generation  tools for Automated Classifcation" was published in Microscopy and Microanalysis 11 on Aug 4, 2005. OR you can get it here


Purdue submitted a provisional patent on April 8, 2004 and the patent US7280204 was issued on Oct 9, 2007. Another paper on our work was published in 2011. This patent covers all applications and use of spectral flow cytometry.


Spectral Signatures


A particular aspect covered by the Purdue patent is the association of spectral signatures to biological activity. This is fundamental to spectral cytometry, so all flow cytometry use of signature creation would infringe the Purdue patent unless it was performed under license (i.e. only by Sony).



The Purdue patent was licensed by Sony Life Sciences and is the basis of the technology used in the Sony Spectral flow cytometer. For some reason SONY has chosen never to publically acknowledge that their spectral flow cytometer is based on a license to the Purdue technology. This is pretty annoying - you would think they invented the idea! ThermoFisher licensed the patent in July 2020.


The issued patent US7280204 covers both the hardware to achieve spectral flow cytometry AND the analytical component. So the use of spectral unmixing for example, to identify a specific probe and define a phenotype is covered under the patent. Only Sony and ThermoFisher have licensed the Purdue patent - any other companies selling flow cytometers for using spectral analysis are infringing the Purdue patent. 


Publications from our Group on Spectral Cytometry

J.Paul Robinson presented the first work on spectral flow cytometry at the 2004 ISAC conference "Collection Hardware for high speed mutlispectral single particle analysis" (see abstract attached). 


Weeks prior to the ISAC conference (April 4, 2004) we submitted a US patent application ("Multispectral detector and analysis system") resulting in # US7,280,204 issued on Oct 9, 2007 with a priority date of April 8, 2004. 


We published a rapid publication in Biophotoncis International , Oct, 2004 "Multispectral Cytometry: The Next Generation" (copy attached) - this was the first paper showing the current implementation of spectral flow cytometry.  


We published a note in Microsc Microanal 11(Suppl 2), 2005 "Multispectral Flow Cytometry: Next Generation tools for Automated Classification" DOI: 10.1017/S1431927605510328 (see attached). 


In addition we published an SPIE paper: (Advanced Biomedical and Clinical Diagnostic Systems III, edited by Tuan Vo-Dinh,
Warren S. Grundfest, David A. Benaron, Gerald E. Cohn, Proc. of SPIE Vol. 5692) entitled "Multispectral cytometry of single bio-particles using a 32-channel
" (attached) . 


Gerald Gregori (then in our group) published a paper in Cytometry 81A:35-44, 2012 "Hyperspectral cytometry at the single cell level using a 32-Channel Photodetector" (attached)


A recent paper in 2019 is here Spectral Flow Cytometry Article published April 30, 2019 or download here


Past Talks on Spectral Flow Cytometry

2006:  Next-Generation Cytomics: Spectral Fingerprinting  (PDF)  Presented at "Biophotonics in Australia: Showcase and Strategic Planning"  held at Macquarie University, Sydney,Australia from 22 to 25 February 2006

2007: "Cytomics and the Integration of Next-Generation Technologies" (PDF)  AFCG Meeting, Melbourne, Australia


2008:  "Advances in Automated Cell Identification and Classification" (PDF) XIII International Congress of Histochemistry and Cytochemistry, Gdansk., Poland


2009:  "Advanced Approaches to flow Analysis" 5th International Advanced Course in Cytometry, University of Modena, March 2009


2010: "Spectral Fingerprinting" KIST-Tohoku Joint Symposium on Nanobiomedical Engineering, Korea (PDF)


2011:  "Historical Perspective of Cytometry: Past successes and Future Opportunities" 3rd Turkish-US Cytometry Wokshop” Akdeniz University, Antalya, Turkey  (PDF)

2011: "Future opportunities in cell analysis" France 2011


2021: "New Frontiers in Flow Cytometry" Iberian Society Meeting, June 2021


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