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.
TY - 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 . .
