Interactions of ultrashort laser pulses with hemoglobin: Photophysical aspects and potential applications
Аутори
Radmilović, Mihajlo D.Drvenica, Ivana
Rabasović, Mihailo D.
Ilić, Vesna
Pavlović, Danica
Oasa, Sho
Vukojević, Vladana
Perić, Mina
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 / Two-photon excitation fluorescence / Hemoglobin photoproduct / Femtosecond laser / Protoporphyrin IXИзвор:
International Journal of Biological Macromolecules, 2023, 244, 125312-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
- Министарство науке, технолошког развоја и иновација Републике Србије, институционално финансирање - 200024 (Универзитет у Београду, Институт за физику, Београд-Земун) (RS-MESTD-inst-2020-200024)
- Министарство науке, технолошког развоја и иновација Републике Србије, институционално финансирање - 200015 (Универзитет у Београду, Институт за медицинска истраживања) (RS-MESTD-inst-2020-200015)
Институција/група
Institut za medicinska istraživanjaTY - JOUR AU - Radmilović, Mihajlo D. AU - Drvenica, Ivana AU - Rabasović, Mihailo D. AU - Ilić, Vesna AU - Pavlović, Danica AU - Oasa, Sho AU - Vukojević, Vladana AU - Perić, Mina AU - Nikolić, Stanko N. AU - Krmpot, Aleksandar J. PY - 2023 UR - http://rimi.imi.bg.ac.rs/handle/123456789/1324 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. T2 - International Journal of Biological Macromolecules 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 and Rabasović, Mihailo D. and Ilić, Vesna 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.", journal = "International Journal of Biological Macromolecules, 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., Rabasović, M. D., Ilić, V., 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, 244, 125312. https://doi.org/10.1016/j.ijbiomac.2023.125312
Radmilović MD, Drvenica I, Rabasović MD, Ilić V, 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, Rabasović, Mihailo D., Ilić, Vesna, 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 . .