Engineered materials induce immune responses when implanted in vivo. The autofluorescence spectra of responding cells can be captured by spectral cytometry. We discuss the necessary considerations when performing spectral cytometry on highly autofluorescent samples to extract phenotypic information from autofluorescence spectra and perform accurate quantification of fluorescent labels.
References
Nolan, J. P. & Condello, D. Spectral flow cytometry. Curr. Protoc. Cytom. https://doi.org/10.1002/0471142956.cy0127s63 (2013).
Adan, A., Alizada, G., Kiraz, Y., Baran, Y. & Nalbant, A. Flow cytometry: basic principles and applications. Crit. Rev. Biotechnol. 37, 163–176 (2017).
Adusei, K. M. et al. Development of a high-color flow cytometry panel for immunologic analysis of tissue injury and reconstruction in a rat model. Cells Tissues Organs https://doi.org/10.1159/000524682 (2022).
Stewart, J. C., Villasmil, M. L. & Frampton, M. W. Changes in fluorescence intensity of selected leukocyte surface markers following fixation. Cytometry A 71, 379–385 (2007).
Saeys, Y., Van Gassen, S. & Lambrecht, B. N. Computational flow cytometry: helping to make sense of high-dimensional immunology data. Nat. Rev. Immunol. 16, 449–462 (2016).
Acknowledgements
This work was funded by the Intramural Research Program of the National Institute for Biomedical Imaging and Bioengineering, and the National Cancer Institute, CCR, CIL of the National Institutes of Health, Bethesda, MD, USA.
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Lokwani, R., Chaudhari, R., Wolf, M.T. et al. Spectral cytometry on highly autofluorescent samples. Nat Rev Methods Primers 2, 71 (2022). https://doi.org/10.1038/s43586-022-00156-0
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DOI: https://doi.org/10.1038/s43586-022-00156-0
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