TY  - JOUR
AU  - Bukara, Katarina
AU  - Jovanić, Svetlana
AU  - Drvenica, Ivana
AU  - Stančić, Ana
AU  - Ilić, Vesna Lj.
AU  - Rabasović, Mihailo D.
AU  - Pantelić, Dejan
AU  - Jelenković, Branislav
AU  - Bugarski, Branko
AU  - Krmpot, Aleksandar
PY  - 2017
UR  - http://TechnoRep.tmf.bg.ac.rs/handle/123456789/3747
AB  - The present study describes utilization of two photon excitation fluorescence (2PE) microscopy for visualization of the hemoglobin in human and porcine erythrocytes and their empty membranes (i.e., ghosts). High-quality, label-and fixation-free visualization of hemoglobin was achieved at excitation wavelength 730 nm by detecting visible autofluorescence. Localization in the suspension and spatial distribution (i.e., mapping) of residual hemoglobin in erythrocyte ghosts has been resolved by 2PE. Prior to the 2PE mapping, the presence of residual hemoglobin in the bulk suspension of erythrocyte ghosts was confirmed by cyanmethemoglobin assay. 2PE analysis revealed that the distribution of hemoglobin in intact erythrocytes follows the cells' shape. Two types of erythrocytes, human and porcine, characterized with discocyte and echinocyte morphology, respectively, showed significant differences in hemoglobin distribution. The 2PE images have revealed that despite an extensive washing out procedure after gradual hypotonic hemolysis, a certain amount of hemoglobin localized on the intracellular side always remains bound to the membrane and cannot be eliminated. The obtained results open the possibility to use 2PE microscopy to examine hemoglobin distribution in erythrocytes and estimate the purity level of erythrocyte ghosts in biotechnological processes.
PB  - Spie-Soc Photo-Optical Instrumentation Engineers, Bellingham
T2  - Journal of Biomedical Optics
T1  - Mapping of hemoglobin in erythrocytes and erythrocyte ghosts using two photon excitation fluorescence microscopy
IS  - 2
VL  - 22
DO  - 10.1117/1.JBO.22.2.026003
ER  - 

@article{
author = "Bukara, Katarina and Jovanić, Svetlana and Drvenica, Ivana and Stančić, Ana and Ilić, Vesna Lj. and Rabasović, Mihailo D. and Pantelić, Dejan and Jelenković, Branislav and Bugarski, Branko and Krmpot, Aleksandar",
year = "2017",
abstract = "The present study describes utilization of two photon excitation fluorescence (2PE) microscopy for visualization of the hemoglobin in human and porcine erythrocytes and their empty membranes (i.e., ghosts). High-quality, label-and fixation-free visualization of hemoglobin was achieved at excitation wavelength 730 nm by detecting visible autofluorescence. Localization in the suspension and spatial distribution (i.e., mapping) of residual hemoglobin in erythrocyte ghosts has been resolved by 2PE. Prior to the 2PE mapping, the presence of residual hemoglobin in the bulk suspension of erythrocyte ghosts was confirmed by cyanmethemoglobin assay. 2PE analysis revealed that the distribution of hemoglobin in intact erythrocytes follows the cells' shape. Two types of erythrocytes, human and porcine, characterized with discocyte and echinocyte morphology, respectively, showed significant differences in hemoglobin distribution. The 2PE images have revealed that despite an extensive washing out procedure after gradual hypotonic hemolysis, a certain amount of hemoglobin localized on the intracellular side always remains bound to the membrane and cannot be eliminated. The obtained results open the possibility to use 2PE microscopy to examine hemoglobin distribution in erythrocytes and estimate the purity level of erythrocyte ghosts in biotechnological processes.",
publisher = "Spie-Soc Photo-Optical Instrumentation Engineers, Bellingham",
journal = "Journal of Biomedical Optics",
title = "Mapping of hemoglobin in erythrocytes and erythrocyte ghosts using two photon excitation fluorescence microscopy",
number = "2",
volume = "22",
doi = "10.1117/1.JBO.22.2.026003"
}

Bukara, K., Jovanić, S., Drvenica, I., Stančić, A., Ilić, V. Lj., Rabasović, M. D., Pantelić, D., Jelenković, B., Bugarski, B.,& Krmpot, A.. (2017). Mapping of hemoglobin in erythrocytes and erythrocyte ghosts using two photon excitation fluorescence microscopy. in Journal of Biomedical Optics
Spie-Soc Photo-Optical Instrumentation Engineers, Bellingham., 22(2).
https://doi.org/10.1117/1.JBO.22.2.026003

Bukara K, Jovanić S, Drvenica I, Stančić A, Ilić VL, Rabasović MD, Pantelić D, Jelenković B, Bugarski B, Krmpot A. Mapping of hemoglobin in erythrocytes and erythrocyte ghosts using two photon excitation fluorescence microscopy. in Journal of Biomedical Optics. 2017;22(2).
doi:10.1117/1.JBO.22.2.026003 .

Bukara, Katarina, Jovanić, Svetlana, Drvenica, Ivana, Stančić, Ana, Ilić, Vesna Lj., Rabasović, Mihailo D., Pantelić, Dejan, Jelenković, Branislav, Bugarski, Branko, Krmpot, Aleksandar, "Mapping of hemoglobin in erythrocytes and erythrocyte ghosts using two photon excitation fluorescence microscopy" in Journal of Biomedical Optics, 22, no. 2 (2017),
https://doi.org/10.1117/1.JBO.22.2.026003 . .