Unlocking autofluorescence in the era of full spectrum analysis: Implications for immunophenotype discovery projects

VJ Jameson; T Luke; Y Yan; A Hind; M Evrard; K Man; LK Mackay; A Kallies; JA Villadangos; HEG McWilliam; A Perez-Gonzalez

Journal article

Unlocking autofluorescence in the era of full spectrum analysis: Implications for immunophenotype discovery projects

VJ Jameson, T Luke, Y Yan, A Hind, M Evrard, K Man, LK Mackay, A Kallies, JA Villadangos, HEG McWilliam, A Perez-Gonzalez

Cytometry Part A | Published : 2022

DOI: 10.1002/cyto.a.24555

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Abstract

Understanding the complex elements affecting signal resolution in cytometry is key for quality experimental design and data. In this study, we incorporate autofluorescence as a contributing factor to our understanding of resolution in cytometry and corroborate its impact in fluorescence signal detection through mathematical predictions supported by empirical evidence. Our findings illustrate the critical importance of autofluorescence extraction via full spectrum unmixing in unmasking dim signals and delineating the expression and subset distribution of low abundance markers in discovery projects. We apply our findings to the precise definition of the tissue and cellular distribution of a we..

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University of Melbourne Researchers

Maximilien Evrard Author

Kevin Man Author

Laura Mackay Author

Axel Kallies Author

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Grants

Awarded by National Institutes of Health

Funding Acknowledgements

Australian Research Council (ARC), Grant/Award Number: DP170102471; CASS Foundation Medicine/Science Grant; Howard Hughes Medical Institute and Bill & Melinda Gates International Research Scholarship, Grant/Award Number: OPP1175796; National Health and Medical Research Council of Australia (NHMRC), Grant/Award Number: 1113293; National Institute of Allergy and Infectious Diseases of the National Institutes of Health (NIH), Grant/Award Number: R01AI148407; NHMRC CJ Martin Fellowship, Grant/Award Number: GNT11219; NHMRC Ideas Grant, Grant/Award Number: 2003192; NHMRC Principal Research Fellowship, Grant/Award Number: 1154592; Phenomics Australia and the Australian Government through the National Collaborative Research Infrastructure Strategy (NCRIS) program; University of Melbourne McKenzie Postdoctoral Fellowship