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dc.creatorBukara, Katarina
dc.creatorJovanić, Svetlana
dc.creatorDrvenica, Ivana
dc.creatorStančić, Ana
dc.creatorIlić, Vesna Lj.
dc.creatorRabasović, Mihailo D.
dc.creatorPantelić, Dejan
dc.creatorJelenković, Branislav
dc.creatorBugarski, Branko
dc.creatorKrmpot, Aleksandar
dc.date.accessioned2021-03-10T13:34:28Z
dc.date.available2021-03-10T13:34:28Z
dc.date.issued2017
dc.identifier.issn1083-3668
dc.identifier.urihttp://TechnoRep.tmf.bg.ac.rs/handle/123456789/3747
dc.description.abstractThe 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.en
dc.publisherSpie-Soc Photo-Optical Instrumentation Engineers, Bellingham
dc.relationQatar National Research Fund (a member of the Qatar Foundation) [NPRP 6-021-1-005]
dc.relationinfo:eu-repo/grantAgreement/MESTD/Integrated and Interdisciplinary Research (IIR or III)/46010/RS//
dc.relationinfo:eu-repo/grantAgreement/MESTD/Basic Research (BR or ON)/171005/RS//
dc.relationinfo:eu-repo/grantAgreement/MESTD/Basic Research (BR or ON)/171038/RS//
dc.relationinfo:eu-repo/grantAgreement/MESTD/Integrated and Interdisciplinary Research (IIR or III)/45016/RS//
dc.rightsopenAccess
dc.rights.urihttps://creativecommons.org/licenses/by/4.0/
dc.sourceJournal of Biomedical Optics
dc.subjectmultiphoton fluorescence microscopyen
dc.subjectultrafast lasersen
dc.subjecthemoglobinen
dc.subjectlabel-free imagingen
dc.subjecterythrocytesen
dc.subjecterythrocyte ghostsen
dc.titleMapping of hemoglobin in erythrocytes and erythrocyte ghosts using two photon excitation fluorescence microscopyen
dc.typearticle
dc.rights.licenseBY
dc.citation.issue2
dc.citation.other22(2): -
dc.citation.rankM22
dc.citation.volume22
dc.identifier.doi10.1117/1.JBO.22.2.026003
dc.identifier.fulltexthttp://TechnoRep.tmf.bg.ac.rs/bitstream/id/9583/026003_1.pdf
dc.identifier.pmid28301654
dc.identifier.scopus2-s2.0-85013159159
dc.identifier.wos000397267900011
dc.type.versionpublishedVersion


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