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Article

Phasor-Based Endogenous NAD(P)H Fluorescence Lifetime Imaging Unravels Specific Enzymatic Activity of Neutrophil Granulocytes Preceding NETosis

1
Biophysical Analytics, German Rheumatism Research Center, 10117 Berlin, Germany
2
Neuropathology, Charité—Universitätsmedizin, 10117 Berlin, Germany
3
Intravital Microscopy and Immune Dynamics, Charité—Universitätsmedizin, 10117 Berlin, Germany
4
Immune Dynamics, German Rheumatism Research Center, 10117 Berlin, Germany
*
Author to whom correspondence should be addressed.
Int. J. Mol. Sci. 2018, 19(4), 1018; https://doi.org/10.3390/ijms19041018
Received: 30 January 2018 / Revised: 15 March 2018 / Accepted: 17 March 2018 / Published: 29 March 2018
(This article belongs to the Special Issue Laser Application in Life Sciences 2018)
Time-correlated single-photon counting combined with multi-photon laser scanning microscopy has proven to be a versatile tool to perform fluorescence lifetime imaging in biological samples and, thus, shed light on cellular functions, both in vitro and in vivo. Here, by means of phasor-analyzed endogenous NAD(P)H (nicotinamide adenine dinucleotide (phosphate)) fluorescence lifetime imaging, we visualize the shift in the cellular metabolism of healthy human neutrophil granulocytes during phagocytosis of Staphylococcus aureus pHrodo™ beads. We correlate this with the process of NETosis, i.e., trapping of pathogens by DNA networks. Hence, we are able to directly show the dynamics of NADPH oxidase activation and its requirement in triggering NETosis in contrast to other pathways of cell death and to decipher the dedicated spatio-temporal sequence between NADPH oxidase activation, nuclear membrane disintegration and DNA network formation. The endogenous FLIM approach presented here uniquely meets the increasing need in the field of immunology to monitor cellular metabolism as a basic mechanism of cellular and tissue functions. View Full-Text
Keywords: NAD(P)H (nicotinamide adenine dinucleotide (phosphate)); fluorescence lifetime imaging; neutrophil granulocytes; NADPH oxidase; NETosis NAD(P)H (nicotinamide adenine dinucleotide (phosphate)); fluorescence lifetime imaging; neutrophil granulocytes; NADPH oxidase; NETosis
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MDPI and ACS Style

Leben, R.; Ostendorf, L.; Van Koppen, S.; Rakhymzhan, A.; Hauser, A.E.; Radbruch, H.; Niesner, R.A. Phasor-Based Endogenous NAD(P)H Fluorescence Lifetime Imaging Unravels Specific Enzymatic Activity of Neutrophil Granulocytes Preceding NETosis. Int. J. Mol. Sci. 2018, 19, 1018. https://doi.org/10.3390/ijms19041018

AMA Style

Leben R, Ostendorf L, Van Koppen S, Rakhymzhan A, Hauser AE, Radbruch H, Niesner RA. Phasor-Based Endogenous NAD(P)H Fluorescence Lifetime Imaging Unravels Specific Enzymatic Activity of Neutrophil Granulocytes Preceding NETosis. International Journal of Molecular Sciences. 2018; 19(4):1018. https://doi.org/10.3390/ijms19041018

Chicago/Turabian Style

Leben, Ruth, Lennard Ostendorf, Sofie Van Koppen, Asylkhan Rakhymzhan, Anja E. Hauser, Helena Radbruch, and Raluca A. Niesner. 2018. "Phasor-Based Endogenous NAD(P)H Fluorescence Lifetime Imaging Unravels Specific Enzymatic Activity of Neutrophil Granulocytes Preceding NETosis" International Journal of Molecular Sciences 19, no. 4: 1018. https://doi.org/10.3390/ijms19041018

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