Összes szerző

Mészáros Mária

az alábbi absztraktok szerzői között szerepel:

Fekete Tamás
Measuring binding force between glutathione-functionalized microtools and blood brain barrier cells with optical tweezer

Aug 28 - szerda

13:30 – 15:30

II. Poszterszekció

P48

Measuring binding force between glutathione-functionalized microtools and blood brain barrier cells with optical tweezer

Tamás Fekete, Mária Mészáros, Gergő Porkoláb, Szilvia Veszelka, Mária Deli, Lóránd Kelemen

Institute of Biophysics, Biological Research Centre of the Hungarian Academy of Sciences, Temesvári krt. 62, H-6726 Szeged, Hungary

In this work functionalized and optically actuated microstructures are used to study the adhesion between cell membrane-associated proteins and their ligands. Literature indicates that such proteins from the solute carrier transporter family can be effectively used in the active transport of drugs to the brain through cells forming the blood brain barrier (BBB). Mészáros and co-workers found if transporter protein ligands, such as alanine, D-glucose or glutathione are mounted on the surface of niosomes encapsulating the drug to be delivered, the uptake rate into the BBB cells increases (Mészáros et al. 2018). In this study we characterize ligand binding by measuring the force between these ligands, immobilized covalently on the optically actuated microstructures and the BBB’s component cells. The complex-shaped microstructures (Aekbote et al 2016) were made by two-photon polymerization and equipped with a flat end of variable surface area that interacts with the cell membrane. The ligand-coated structures are pressed against the cells, grown on a vertical support, and when retracted, the rupture force is measured. Since optical tweezers can exert lower forces that an AFM cantilever, these experiments can elucidate finer interactions responsible for the ligand binding to the BBB cells surface.

References

Mészáros M, Porkoláb G, Kiss L et al. Niosomes decorated with dual ligands targeting brain endothelial transporters increase cargo penetration across the blood-brain barrier, Eur J Pharm Sci, 123:228–240, 2018, doi.org/10.1016/j.ejps.2018.07.042

Aekbote BL, Fekete T, Jacak J, Vizsnyiczai G, Ormos P, and Kelemen L, Surface-modified complex SU-8 microstructures for indirect optical manipulation of single cells, Biomed Optics Express, 7:45-56, 2016, doi.org/10.1364/BOE.7.000045

Vizsnyiczai Gaszton
Don’t be a fool, use a microtool: Biophotonic toolbox for single cell studies

Aug 28 - szerda

13:30 – 15:30

II. Poszterszekció

P52

Don’t be a fool, use a microtool: Biophotonic toolbox for single cell studies 

Gaszton Vizsnyiczai¹, Tamás Fekete¹, Mária Mészáros¹, András Búzás¹, Gergely Iványi¹, Ádám Apró¹, Pál Ormos¹, Rebeca Martínez Vázquez² and Lóránd Kelemen¹

¹ Biological Research Centre, Institute of Biophysics, Hungarian Academy of Sciences, Temesvári krt. 62, 6726 Szeged, Hungary

² Institute for Photonics and Nanotechnologies, National Research Council, Piazza Leonardo da Vinci, 32, 20133 Milan, Italy

Microfluidics has revolutionized biological research due to its capabilities that enables measurements in extremely small volumes in a highly parallelized and integrated manner, for a relatively low cost. Considering cell studies in a microfluidic system, engineered microenvironments can add extra possibilities to the experiments: besides the plain walls, geometrical or chemical modifications, or even microdevices can be created inside the microfluidic channels. Here we introduce an assortment of microstructures and tools we have created, tested and offer for single cell studies in microchannels. These include channel-attached as well as mobile devices which can be actuated with optical tweezers, as follows.

We fabricated microslits with sub-micrometer opening to study cancer cell migration through confined spaces.

An integrated whispering-gallery mode detector was made inside a microchannel and the presence of protein was detected with it in micromolar concentration in a selective way.

The overall deformability of a single cell can be measured with a two-arm micro lever that can multiply the force the optical tweezers exerts.

Local cell deformability and its Young-modulus were obtained with an optically actuated microtool that can make small indentation on the cell membrane.

A similar tool with functionalized surface is used to test adhesion forces of various ligands to their corresponding membrane-bound receptors.

Multiview microscopic imaging of single cell was performed using microtools that are attached to the cells enabling their indirect optical manipulation; in cases when attachment is not possible, two manipulators can be used to grab and move a cell.

We plan to extend our fluorescent microscopic system which is outfitted with the optical trap towards light sheet excitation to improve image equality and data collection speed. One of our planned biological applications is to study single cell interactions between various fungi species and immune cells.

Acknowledgements

This work was supported by funding received from the CONCERT-Japan Photonic Manufacturing Joint Call (FEASIBLE project), GINOP-2.3.2-15- 2016-00001, GINOP-2.3.3-15-2016-00040 and the European Union's Horizon 2020 Research and Innovation Programme under grant agreement No. 654148 Laserlab-Europe.