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Interactions of ultrashort laser pulses with hemoglobin: Photophysical aspects and potential applications

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submitted on 2023-07-13, 05:14 and posted on 2023-07-16, 12:20 authored by Mihajlo D. Radmilović, Ivana T. Drvenica, Mihailo D. Rabasović, Vesna Lj. Ilić, Danica Pavlović, Sho Oasa, Vladana Vukojević, Mina Perić, Stanko N. Nikolić, Aleksandar J. Krmpot

Hemoglobin (Hb), a life-sustaining and highly abundant erythrocyte protein, is not readily fluorescent. A few studies have already reported Two-Photon Excited Fluorescence (TPEF) of Hb, however, the mechanisms through which Hb becomes fluorescent upon interaction with ultrashort laser pulses are not completely understood. Here, we characterized photophysically this interaction on Hb thin film and erythrocytes using fluorescence spectroscopy upon single-photon/two-photon absorption, and UV-VIS single-photon absorption spectroscopy. A gradual increase of the fluorescence intensity, ending up with saturation, is observed upon prolonged exposure of Hb thin layer and erythrocytes to ultrashort laser pulses at 730 nm. When compared to protoporphyrin IX (PpIX) and oxidized Hb by H2O2, TPEF spectra from a thin Hb film and erythrocytes showed good mutual agreement, broad peaking at 550 nm, supporting hemoglobin undergoes degradation and that same fluorescent specie(s) originating from the heme moiety are generated. The uniform square shaped patterns of the fluorescent photoproduct exhibited the same level of the fluorescence intensity even after 12 weeks from the formation, indicating high photoproduct stability. We finally demonstrated the full potential of the formed Hb photoproduct with TPEF scanning microscopy towards spatiotemporally controlled micropatterning in HTF and single human erythrocyte labelling and tracking in the whole blood.  

Other Information

Published in: International Journal of Biological Macromolecules
License: https://creativecommons.org/licenses/by/4.0/
See article on publisher's website: http://dx.doi.org/10.1016/j.ijbiomac.2023.125312 

Funding

Open Access funding provided by the Qatar National Library

History

Language

  • English

Publisher

Elsevier

Publication Year

  • 2023

License statement

This Item is licensed under the Creative Commons Attribution 4.0 International License

Institution affiliated with

  • Texas A&M University at Qatar