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

<p>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 pulse...

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Main Author: Mihajlo D. Radmilović (16538659) (author)
Other Authors: Ivana T. Drvenica (16538660) (author), Mihailo D. Rabasović (16538661) (author), Vesna Lj. Ilić (16538664) (author), Danica Pavlović (16538667) (author), Sho Oasa (7244462) (author), Vladana Vukojević (165297) (author), Mina Perić (8859218) (author), Stanko N. Nikolić (16538668) (author), Aleksandar J. Krmpot (16538671) (author)
Published: 2023
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author Mihajlo D. Radmilović (16538659)
author2 Ivana T. Drvenica (16538660)
Mihailo D. Rabasović (16538661)
Vesna Lj. Ilić (16538664)
Danica Pavlović (16538667)
Sho Oasa (7244462)
Vladana Vukojević (165297)
Mina Perić (8859218)
Stanko N. Nikolić (16538668)
Aleksandar J. Krmpot (16538671)
author2_role author
author
author
author
author
author
author
author
author
author_facet Mihajlo D. Radmilović (16538659)
Ivana T. Drvenica (16538660)
Mihailo D. Rabasović (16538661)
Vesna Lj. Ilić (16538664)
Danica Pavlović (16538667)
Sho Oasa (7244462)
Vladana Vukojević (165297)
Mina Perić (8859218)
Stanko N. Nikolić (16538668)
Aleksandar J. Krmpot (16538671)
author_role author
dc.creator.none.fl_str_mv Mihajlo D. Radmilović (16538659)
Ivana T. Drvenica (16538660)
Mihailo D. Rabasović (16538661)
Vesna Lj. Ilić (16538664)
Danica Pavlović (16538667)
Sho Oasa (7244462)
Vladana Vukojević (165297)
Mina Perić (8859218)
Stanko N. Nikolić (16538668)
Aleksandar J. Krmpot (16538671)
dc.date.none.fl_str_mv 2023-07-31T00:00:00Z
dc.identifier.none.fl_str_mv 10.1016/j.ijbiomac.2023.125312
dc.relation.none.fl_str_mv https://figshare.com/articles/journal_contribution/Interactions_of_ultrashort_laser_pulses_with_hemoglobin_Photophysical_aspects_and_potential_applications/23674215
dc.rights.none.fl_str_mv CC BY 4.0
info:eu-repo/semantics/openAccess
dc.subject.none.fl_str_mv Biological sciences
Biochemistry and cell biology
Biomedical and clinical sciences
Cardiovascular medicine and haematology
Erythrocytes
Two-photon excitation fluorescence
Hemoglobin photoproduct
Femtosecond laser
Protoporphyrin IX
dc.title.none.fl_str_mv Interactions of ultrashort laser pulses with hemoglobin: Photophysical aspects and potential applications
dc.type.none.fl_str_mv Text
Journal contribution
info:eu-repo/semantics/publishedVersion
text
contribution to journal
description <p>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 H<sub>2</sub>O<sub>2</sub>, 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.  </p> <h2>Other Information</h2> <p>Published in: International Journal of Biological Macromolecules<br> License: <a href="https://creativecommons.org/licenses/by/4.0/" target="_blank">https://creativecommons.org/licenses/by/4.0/</a><br> See article on publisher's website: <a href="http://dx.doi.org/10.1016/j.ijbiomac.2023.125312" target="_blank">http://dx.doi.org/10.1016/j.ijbiomac.2023.125312 </a></p>
eu_rights_str_mv openAccess
id Manara2_c541522e2b228040cb35e9b568cab0f2
identifier_str_mv 10.1016/j.ijbiomac.2023.125312
network_acronym_str Manara2
network_name_str Manara2
oai_identifier_str oai:figshare.com:article/23674215
publishDate 2023
repository.mail.fl_str_mv
repository.name.fl_str_mv
repository_id_str
rights_invalid_str_mv CC BY 4.0
spelling Interactions of ultrashort laser pulses with hemoglobin: Photophysical aspects and potential applicationsMihajlo D. Radmilović (16538659)Ivana T. Drvenica (16538660)Mihailo D. Rabasović (16538661)Vesna Lj. Ilić (16538664)Danica Pavlović (16538667)Sho Oasa (7244462)Vladana Vukojević (165297)Mina Perić (8859218)Stanko N. Nikolić (16538668)Aleksandar J. Krmpot (16538671)Biological sciencesBiochemistry and cell biologyBiomedical and clinical sciencesCardiovascular medicine and haematologyErythrocytesTwo-photon excitation fluorescenceHemoglobin photoproductFemtosecond laserProtoporphyrin IX<p>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 H<sub>2</sub>O<sub>2</sub>, 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.  </p> <h2>Other Information</h2> <p>Published in: International Journal of Biological Macromolecules<br> License: <a href="https://creativecommons.org/licenses/by/4.0/" target="_blank">https://creativecommons.org/licenses/by/4.0/</a><br> See article on publisher's website: <a href="http://dx.doi.org/10.1016/j.ijbiomac.2023.125312" target="_blank">http://dx.doi.org/10.1016/j.ijbiomac.2023.125312 </a></p>2023-07-31T00:00:00ZTextJournal contributioninfo:eu-repo/semantics/publishedVersiontextcontribution to journal10.1016/j.ijbiomac.2023.125312https://figshare.com/articles/journal_contribution/Interactions_of_ultrashort_laser_pulses_with_hemoglobin_Photophysical_aspects_and_potential_applications/23674215CC BY 4.0info:eu-repo/semantics/openAccessoai:figshare.com:article/236742152023-07-31T00:00:00Z
spellingShingle Interactions of ultrashort laser pulses with hemoglobin: Photophysical aspects and potential applications
Mihajlo D. Radmilović (16538659)
Biological sciences
Biochemistry and cell biology
Biomedical and clinical sciences
Cardiovascular medicine and haematology
Erythrocytes
Two-photon excitation fluorescence
Hemoglobin photoproduct
Femtosecond laser
Protoporphyrin IX
status_str publishedVersion
title Interactions of ultrashort laser pulses with hemoglobin: Photophysical aspects and potential applications
title_full Interactions of ultrashort laser pulses with hemoglobin: Photophysical aspects and potential applications
title_fullStr Interactions of ultrashort laser pulses with hemoglobin: Photophysical aspects and potential applications
title_full_unstemmed Interactions of ultrashort laser pulses with hemoglobin: Photophysical aspects and potential applications
title_short Interactions of ultrashort laser pulses with hemoglobin: Photophysical aspects and potential applications
title_sort Interactions of ultrashort laser pulses with hemoglobin: Photophysical aspects and potential applications
topic Biological sciences
Biochemistry and cell biology
Biomedical and clinical sciences
Cardiovascular medicine and haematology
Erythrocytes
Two-photon excitation fluorescence
Hemoglobin photoproduct
Femtosecond laser
Protoporphyrin IX