Interactions of ultrashort laser pulses with hemoglobin: Photophysical aspects and potential applications
Аутори
Radmilović, Mihajlo D.Drvenica, Ivana T.
Rabasović, Mihailo D.
Ilić, Vesna Lj.
Pavlović, Danica
Oasa, Sho
Vukojević, Vladana
Perić, Mina
![](/themes/Mirageimagine/images/orcid.png)
Nikolić, Stanko N.
Krmpot, Aleksandar J.
Чланак у часопису (Објављена верзија)
Метаподаци
Приказ свих података о документуАпстракт
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.
Кључне речи:
Erythrocytes / Femtosecond laser / Hemoglobin photoproduct / Protoporphyrin IX / Two-photon excitation fluorescenceИзвор:
International Journal of Biological Macromolecules, 2023, 244, 125312-Издавач:
- Elsevier
Финансирање / пројекти:
- HEMMAGINERO - Hemoglobin-Based Spectroscopy and Nonlinear Imaging of Erythrocytes and Their Membranes As Emerging Diagnostic Tool (RS-ScienceFundRS-Promis-6066079)
- Qatar National Research Fund [grant number PPM 04-0131-200019]
URI
https://www.sciencedirect.com/science/article/pii/S0141813023022067https://imagine.imgge.bg.ac.rs/handle/123456789/2288
Институција/група
Institut za molekularnu genetiku i genetičko inženjerstvoTY - JOUR AU - Radmilović, Mihajlo D. AU - Drvenica, Ivana T. AU - Rabasović, Mihailo D. AU - Ilić, Vesna Lj. AU - Pavlović, Danica AU - Oasa, Sho AU - Vukojević, Vladana AU - Perić, Mina AU - Nikolić, Stanko N. AU - Krmpot, Aleksandar J. PY - 2023 UR - https://www.sciencedirect.com/science/article/pii/S0141813023022067 UR - https://imagine.imgge.bg.ac.rs/handle/123456789/2288 AB - 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. PB - Elsevier T2 - International Journal of Biological Macromolecules T1 - Interactions of ultrashort laser pulses with hemoglobin: Photophysical aspects and potential applications SP - 125312 VL - 244 DO - 10.1016/j.ijbiomac.2023.125312 ER -
@article{ author = "Radmilović, Mihajlo D. and Drvenica, Ivana T. and Rabasović, Mihailo D. and Ilić, Vesna Lj. and Pavlović, Danica and Oasa, Sho and Vukojević, Vladana and Perić, Mina and Nikolić, Stanko N. and Krmpot, Aleksandar J.", year = "2023", abstract = "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.", publisher = "Elsevier", journal = "International Journal of Biological Macromolecules", title = "Interactions of ultrashort laser pulses with hemoglobin: Photophysical aspects and potential applications", pages = "125312", volume = "244", doi = "10.1016/j.ijbiomac.2023.125312" }
Radmilović, M. D., Drvenica, I. T., Rabasović, M. D., Ilić, V. Lj., Pavlović, D., Oasa, S., Vukojević, V., Perić, M., Nikolić, S. N.,& Krmpot, A. J.. (2023). Interactions of ultrashort laser pulses with hemoglobin: Photophysical aspects and potential applications. in International Journal of Biological Macromolecules Elsevier., 244, 125312. https://doi.org/10.1016/j.ijbiomac.2023.125312
Radmilović MD, Drvenica IT, Rabasović MD, Ilić VL, Pavlović D, Oasa S, Vukojević V, Perić M, Nikolić SN, Krmpot AJ. Interactions of ultrashort laser pulses with hemoglobin: Photophysical aspects and potential applications. in International Journal of Biological Macromolecules. 2023;244:125312. doi:10.1016/j.ijbiomac.2023.125312 .
Radmilović, Mihajlo D., Drvenica, Ivana T., Rabasović, Mihailo D., Ilić, Vesna Lj., Pavlović, Danica, Oasa, Sho, Vukojević, Vladana, Perić, Mina, Nikolić, Stanko N., Krmpot, Aleksandar J., "Interactions of ultrashort laser pulses with hemoglobin: Photophysical aspects and potential applications" in International Journal of Biological Macromolecules, 244 (2023):125312, https://doi.org/10.1016/j.ijbiomac.2023.125312 . .