Multicolor lifetime imaging and its application to HIV-1 uptake

Tobias Starling, Irene Carlon-Andres, Maro Iliopoulou, Benedikt Kraemer, Maria Loidolt-Krueger, David J. Williamson and Sergi Padilla-Parra ()
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Tobias Starling: Faculty of Life Sciences & Medicine
Irene Carlon-Andres: Faculty of Life Sciences & Medicine
Maro Iliopoulou: University of Oxford
Benedikt Kraemer: Rudower Chaussee 29 (IGZ)
Maria Loidolt-Krueger: Rudower Chaussee 29 (IGZ)
David J. Williamson: Faculty of Life Sciences & Medicine
Sergi Padilla-Parra: Faculty of Life Sciences & Medicine

Nature Communications, 2023, vol. 14, issue 1, 1-17

Abstract: Abstract Simultaneous imaging of nine fluorescent proteins is demonstrated in a single acquisition using fluorescence lifetime imaging microscopy combined with pulsed interleaved excitation of three laser lines. Multicolor imaging employing genetically encodable fluorescent proteins permits spatio-temporal live cell imaging of multiple cues. Here, we show that multicolor lifetime imaging allows visualization of quadruple labelled human immunodeficiency viruses on host cells that in turn are also labelled with genetically encodable fluorescent proteins. This strategy permits to simultaneously visualize different sub-cellular organelles (mitochondria, cytoskeleton, and nucleus) during the process of virus entry with the potential of imaging up to nine different spectral channels in living cells.

Date: 2023
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DOI: 10.1038/s41467-023-40731-x

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