TY  - JOUR
AU  - Pajic, Tanja
AU  - Stevanović, Katarina
AU  - Todorović, Nataša
AU  - Krmpot, Aleksandar J.
AU  - Živić, Miroslav
AU  - Savić-Šević, Svetlana
AU  - Lević, Steva M.
AU  - Stanić, Marina
AU  - Pantelić, Dejan
AU  - Jelenković, Brana
AU  - Rabasović, Mihailo
PY  - 2024
UR  - http://rimsi.imsi.bg.ac.rs/handle/123456789/3213
AB  - Studying the membrane physiology of filamentous fungi is key to understanding their interactions with the environment and crucial for developing new therapeutic strategies for disease-causing pathogens. However, their plasma membrane has been inaccessible for a micron-sized patch-clamp pipette for pA current recordings due to the rigid chitinous cell wall. Here, we report the first femtosecond IR laser nanosurgery of the cell wall of the filamentous fungi, which enabled patch-clamp measurements on protoplasts released from hyphae. A reproducible and highly precise (diffraction-limited, submicron resolution) method for obtaining viable released protoplasts was developed. Protoplast release from the nanosurgery-generated incisions in the cell wall was achieved from different regions of the hyphae. The plasma membrane of the obtained protoplasts formed tight and high-resistance (GΩ) contacts with the recording pipette. The entire nanosurgical procedure followed by the patch-clamp technique could be completed in less than 1 hour. Compared to previous studies using heterologously expressed channels, this technique provides the opportunity to identify new ionic currents and to study the properties of the ion channels in the protoplasts of filamentous fungi in their native environment.
PB  - Springer Nature
T2  - Microsystems & Nanoengineering
T1  - In vivo femtosecond laser nanosurgery of the cell wall enabling patch-clamp measurements on filamentous fungi
SP  - 47
VL  - 10
DO  - 10.1038/s41378-024-00664-x
ER  - 

@article{
author = "Pajic, Tanja and Stevanović, Katarina and Todorović, Nataša and Krmpot, Aleksandar J. and Živić, Miroslav and Savić-Šević, Svetlana and Lević, Steva M. and Stanić, Marina and Pantelić, Dejan and Jelenković, Brana and Rabasović, Mihailo",
year = "2024",
abstract = "Studying the membrane physiology of filamentous fungi is key to understanding their interactions with the environment and crucial for developing new therapeutic strategies for disease-causing pathogens. However, their plasma membrane has been inaccessible for a micron-sized patch-clamp pipette for pA current recordings due to the rigid chitinous cell wall. Here, we report the first femtosecond IR laser nanosurgery of the cell wall of the filamentous fungi, which enabled patch-clamp measurements on protoplasts released from hyphae. A reproducible and highly precise (diffraction-limited, submicron resolution) method for obtaining viable released protoplasts was developed. Protoplast release from the nanosurgery-generated incisions in the cell wall was achieved from different regions of the hyphae. The plasma membrane of the obtained protoplasts formed tight and high-resistance (GΩ) contacts with the recording pipette. The entire nanosurgical procedure followed by the patch-clamp technique could be completed in less than 1 hour. Compared to previous studies using heterologously expressed channels, this technique provides the opportunity to identify new ionic currents and to study the properties of the ion channels in the protoplasts of filamentous fungi in their native environment.",
publisher = "Springer Nature",
journal = "Microsystems & Nanoengineering",
title = "In vivo femtosecond laser nanosurgery of the cell wall enabling patch-clamp measurements on filamentous fungi",
pages = "47",
volume = "10",
doi = "10.1038/s41378-024-00664-x"
}

Pajic, T., Stevanović, K., Todorović, N., Krmpot, A. J., Živić, M., Savić-Šević, S., Lević, S. M., Stanić, M., Pantelić, D., Jelenković, B.,& Rabasović, M.. (2024). In vivo femtosecond laser nanosurgery of the cell wall enabling patch-clamp measurements on filamentous fungi. in Microsystems & Nanoengineering
Springer Nature., 10, 47.
https://doi.org/10.1038/s41378-024-00664-x

Pajic T, Stevanović K, Todorović N, Krmpot AJ, Živić M, Savić-Šević S, Lević SM, Stanić M, Pantelić D, Jelenković B, Rabasović M. In vivo femtosecond laser nanosurgery of the cell wall enabling patch-clamp measurements on filamentous fungi. in Microsystems & Nanoengineering. 2024;10:47.
doi:10.1038/s41378-024-00664-x .

Pajic, Tanja, Stevanović, Katarina, Todorović, Nataša, Krmpot, Aleksandar J., Živić, Miroslav, Savić-Šević, Svetlana, Lević, Steva M., Stanić, Marina, Pantelić, Dejan, Jelenković, Brana, Rabasović, Mihailo, "In vivo femtosecond laser nanosurgery of the cell wall enabling patch-clamp measurements on filamentous fungi" in Microsystems & Nanoengineering, 10 (2024):47,
https://doi.org/10.1038/s41378-024-00664-x . .

TY  - JOUR
AU  - Dmitrović, Svetlana
AU  - Nikolic, Marko G.
AU  - Jelenković, Branislav
AU  - Prekajski, Marija
AU  - Rabasović, Mihailo
AU  - Zarubica, Aleksandra
AU  - Branković, Goran
AU  - Matović, Branko
PY  - 2017
UR  - http://rimsi.imsi.bg.ac.rs/handle/123456789/1056
AB  - Spider dragline silk was coated with pure as well as Eu-doped ceria nanopowders at the room temperature. The treatment was done by immersion of the spider silkmesh into aqueous solutions of cerium nitrate (Ce(NO3)(3)) and ammonium hydroxide (NH4OH). Depending on the relationship between Ce3+ ion and ammonium hydroxide concentration, coated fibers exhibited a different thickness. Obtained materials were studied by means of FESEM. It was found that ceria nanoparticles of average size of 3 nm were coated along spider thread. X-ray diffraction (XRD) and selected-area electron diffraction (SAED) confirmed crystal nature of nanoparticle coating of spider silk. By using Williamson-Hall plots, crystallite size and strain were estimated. EDS measurement confirmed the presence of Eu in spider-Eu-doped ceria composite, and according to FTIR analysis, the interaction between CeO2 and spider silk was proposed. The morphology of obtained composite was observed by TEM. The photoluminescence emission spectra of spider silk coated with Eu-doped ceria were measured with two different excitations of 385 and 466 nm. The two-photon excited auto-fluorescence of spider silk coated with Eu-doped ceria was detected using a nonlinear laser scanning microscope. Obtained composite has a potential as a fluorescent labeling material in diverse applications.
PB  - Springer, Dordrecht
T2  - Journal of Nanoparticle Research
T1  - Photoluminescent properties of spider silk coated with Eu-doped nanoceria
IS  - 2
VL  - 19
DO  - 10.1007/s11051-017-3750-9
ER  - 

@article{
author = "Dmitrović, Svetlana and Nikolic, Marko G. and Jelenković, Branislav and Prekajski, Marija and Rabasović, Mihailo and Zarubica, Aleksandra and Branković, Goran and Matović, Branko",
year = "2017",
abstract = "Spider dragline silk was coated with pure as well as Eu-doped ceria nanopowders at the room temperature. The treatment was done by immersion of the spider silkmesh into aqueous solutions of cerium nitrate (Ce(NO3)(3)) and ammonium hydroxide (NH4OH). Depending on the relationship between Ce3+ ion and ammonium hydroxide concentration, coated fibers exhibited a different thickness. Obtained materials were studied by means of FESEM. It was found that ceria nanoparticles of average size of 3 nm were coated along spider thread. X-ray diffraction (XRD) and selected-area electron diffraction (SAED) confirmed crystal nature of nanoparticle coating of spider silk. By using Williamson-Hall plots, crystallite size and strain were estimated. EDS measurement confirmed the presence of Eu in spider-Eu-doped ceria composite, and according to FTIR analysis, the interaction between CeO2 and spider silk was proposed. The morphology of obtained composite was observed by TEM. The photoluminescence emission spectra of spider silk coated with Eu-doped ceria were measured with two different excitations of 385 and 466 nm. The two-photon excited auto-fluorescence of spider silk coated with Eu-doped ceria was detected using a nonlinear laser scanning microscope. Obtained composite has a potential as a fluorescent labeling material in diverse applications.",
publisher = "Springer, Dordrecht",
journal = "Journal of Nanoparticle Research",
title = "Photoluminescent properties of spider silk coated with Eu-doped nanoceria",
number = "2",
volume = "19",
doi = "10.1007/s11051-017-3750-9"
}

Dmitrović, S., Nikolic, M. G., Jelenković, B., Prekajski, M., Rabasović, M., Zarubica, A., Branković, G.,& Matović, B.. (2017). Photoluminescent properties of spider silk coated with Eu-doped nanoceria. in Journal of Nanoparticle Research
Springer, Dordrecht., 19(2).
https://doi.org/10.1007/s11051-017-3750-9

Dmitrović S, Nikolic MG, Jelenković B, Prekajski M, Rabasović M, Zarubica A, Branković G, Matović B. Photoluminescent properties of spider silk coated with Eu-doped nanoceria. in Journal of Nanoparticle Research. 2017;19(2).
doi:10.1007/s11051-017-3750-9 .

Dmitrović, Svetlana, Nikolic, Marko G., Jelenković, Branislav, Prekajski, Marija, Rabasović, Mihailo, Zarubica, Aleksandra, Branković, Goran, Matović, Branko, "Photoluminescent properties of spider silk coated with Eu-doped nanoceria" in Journal of Nanoparticle Research, 19, no. 2 (2017),
https://doi.org/10.1007/s11051-017-3750-9 . .