I share my opinions on the benefits of and bottlenecks for hyperspectral and time-resolved imaging. I also discuss current and future perspectives for analyzing these types of data using the phasor approach.
References
Balzarotti, F. et al. Science 355, 606–612 (2017).
Digman, M. A., Stakic, M. & Gratton, E. In Fluorescence Fluctuation Spectroscopy (FFS) Part A (ed. Tetin, S. Y.) Ch. 6, 121–144 (Academic, 2013).
Malacrida, L., Ranjit, S., Jameson, D. M. & Gratton, E. Annu. Rev. Biophys. 50, 575–593 (2021).
Torrado, B., Malacrida, L. & Ranjit, S. Sensors (Basel) 22, 999 (2022).
Hanley, Q. S. & Clayton, A. H. A. J. Microsc. 218, 62–67 (2005).
Digman, M. A., Caiolfa, V. R., Zamai, M. & Gratton, E. Biophys. J. 94, L14–L16 (2008).
Redford, G. I. & Clegg, R. M. J. Fluoresc. 15, 805–815 (2005).
Weber, G. J. Phys. Chem. 85, 949–953 (1981).
Jameson, D. M., Gratton, E. & Hall, R. D. Appl. Spectrosc. Rev. 20, 55–106 (1984).
Ranjit, S., Malacrida, L., Jameson, D. M. & Gratton, E. Nat. Protoc. 13, 1979–2004, https://doi.org/10.1038/s41596-018-0026-5 (2018).
Fereidouni, F., Bader, A. N. & Gerritsen, H. C. Opt. Express 20, 12729–12741 (2012).
Sarmento, M. J. et al. Nat. Commun. 9, 3415 (2018).
Vergeldt, F. J. et al. Sci. Rep. 7, 861 (2017).
Ma, N., Digman, M. A., Malacrida, L. & Gratton, E. Biomed. Opt. Express 7, 2441–2452 (2016).
Vu, T. et al. Nat. Commun. 13, 169 (2022).
Vallmitjana, A., Lepanto, P., Irigoin, F. & Malacrida, L. Methods Appl. Fluoresc. 11, 14001 (2023).
Stringari, C., Nourse, J. L., Flanagan, L. A. & Gratton, E. PLoS One 7, e48014 (2012).
Malacrida, L., Jameson, D. M. & Gratton, E. Sci. Rep. 7, 9215 (2017).
Vorontsova, I. et al. Sci. Adv. 8, eabj4833 (2022).
Castello, M. et al. Nat. Methods 16, 175–178 (2019).
Dvornikov, A., Malacrida, L. & Gratton, E. Methods Protoc. 2, 53 (2019).
Hedde, P. N., Cinco, R., Malacrida, L., Kamaid, A. & Gratton, E. Commun. Biol 4, 721 (2021).
Ranjit, S., Malacrida, L. & Gratton, E. Microsc. Res. Tech. 81, 980–989 (2018).
Vallmitjana, A., Torrado, B. & Gratton, E. Biomed. Opt. Express 12, 3410–3422 (2021).
Chiang, H. J. et al. Nat. Methods 20, 248–258 (2023).
Acknowledgements
L.M. is indebted to David M. Jameson for critical reading and valuable suggestions. L.M. is supported by the grants 2020-225439, 2021-240122 and 2022-252604 from the Chan Zuckerberg Initiative DAF, an advised fund of the Silicon Valley Community Foundation; by FOCEM - Fondo para la Convergencia Estructural del Mercosur (COF 03/11); and by Comisión Sectorial de Investigación Científica (CSIC) I+D – 2020 grant #606.
Author information
Authors and Affiliations
Corresponding author
Ethics declarations
Competing Interests
The author declares no competing interests.
Rights and permissions
About this article
Cite this article
Malacrida, L. Phasor plots and the future of spectral and lifetime imaging. Nat Methods 20, 965–967 (2023). https://doi.org/10.1038/s41592-023-01906-y
Published:
Issue Date:
DOI: https://doi.org/10.1038/s41592-023-01906-y
- Springer Nature America, Inc.