MBFT XXIX. Kongresszus - 2023, Budapest .

Összes szerző

Rajmon Imola

az alábbi absztraktok szerzői között szerepel:

Balogh Anna Label-free tracking of cell adhesion kinetics as a function of various parameters

Aug 30 - szerda

15:30 – 17:00

II. Poszterszekció

P35

Label-free tracking of cell adhesion kinetics as a function of various parameters

Anna Balogh^1,2, Kinga Dora Kovacs^1,2, Imola Rajmon^1,2, Inna Szekacs²,

Beatrix Peter², Robert Horvath²

¹Eötvös Lóránd University, Department of Biological Physics, Budapest, Hungary

² Nanobiosensorics Laboratory, Institute of Technical Physics and Materials Science, Centre for Energy Research

Most tissue cells cannot survive for more than a few hours without adherence, if we were able to prevent the adhesion of malignant cells, metastasis could be prevented [1] [2]. The data used for analysis was recorded by a surface sensitive, label-free, resonant waveguide grating based optical biosensor. Due to the force calibration of the optical signal provided by the sensor for individual cells, this technique is suitable for determining the force curves of a large number of cells [3]. To analyse the adhesion signals at the molecular level, we applied and developed different kinetic models. These models take into account relevant molecular parameters, such as dissociation and association rates, the two dimensional kinetic dissociation constant which describes the integrin-ligand binding strength [4].

We investigated the extent to which the adhesion kinetics of cells are affected by the surface density of the cells, the density of echistatin, and the effect of gold nanoparticles on cell adhesion. Echistatin is a potent inhibitor of αIIβ3, αvβ3 and α5β1 receptors [5]. Functionalized nanoparticles can penetrate into living cells, Peter et. al. demonstrated from the recorded kinetic adhesion data that the uptake of the functionalized nanoparticles is an active process [6]. Peter et. al. also investigated the role of glycocalyx components in the cellular uptake of nanoparticles [7]. Subjecting these results to further analysis, we developed procedures and codes that enable the study of molecular parameters influencing adhesion in living cells, in their real environment, without isolating the relevant molecules. Our results potentially open the way for further analysis of the kinetic data obtained from the adhering cells.

Acknowledgment

This work was supported by the National Research, Development, and Innovation Office (Grant Numbers: PD 134195 for Z.Sz, PD 131543 for B.P., ELKH topic-fund, "Élvonal" KKP_19 TKP2022-EGA-04 grants).

References

[1] Sudhakar A. History of Cancer, Ancient and Modern Treatment Methods. J Cancer Sci Ther. 1, 2 (2009). https://doi.org/10.4172/1948-5956.100000e2

[2] Lodish, H., Berk, A., Matsudaira, P., et al. Molecular Cell Biology. (2003)

[3] Sztilkovics, M., Gerecsei, T., Peter, B. et al. Single-cell adhesion force kinetics of cell populations from combined label-free optical biosensor and robotic fluidic force microscopy. Sci Rep 10, 61 (2020). https://doi.org/10.1038/s41598-019-56898-7

[4] Kanyo, N., Kovacs, K.D., Saftics, A. et al. Glycocalyx regulates the strength and kinetics of cancer cell adhesion revealed by biophysical models based on high resolution label-free optical data. Sci Rep 10, 22422 (2020). https://doi.org/10.1038/s41598- 020-80033-6

[5] Szekacs, I., Orgovan, N., Peter, B., et. al. Receptor specific adhesion assay for the quantification of integrin–ligand interactions in intact cells using a microplate based, label-free optical biosensor. Sensors and Actuators B: Chemical, 256 (2018). https://doi.org/10.1016/j.snb.2017.09.208

[6] Peter, B., Lagzi, I., Teraji, S., et. al. Interaction of Positively Charged Gold Nanoparticles with Cancer Cells Monitored by an in Situ Label-Free Optical Biosensor and Transmission Electron Microscopy. ACS Appl. Mater. Interfaces 10, 32 (2018). https://doi.org/10.1021/acsami.8b01546

[7] Peter, B., Kanyo, N., Kovacs, K.D., et. al Glycocalyx Components Detune the Cellular Uptake of Gold Nanoparticles in a Size- and Charge-Dependent Manner. ACS Appl. Bio Mater. 6, 1 (2022). https://doi.org/10.1021/acsabm.2c00595

Rajmon Imola Single-cell adhesion measurements using fluidic force microscopy

Aug 29 - kedd

15:30 – 17:00

I. Poszterszekció

P23

Single-cell adhesion measurements using fluidic force microscopy

Imola Rajmon^1,2, Anna Balogh^1,2, Kinga Dóra Kovács^1,2, Inna Szekács², Robert Horvath²

¹ELTE Eötvös Loránd University, Department of Biological Physics, Budapest, Hungary

²Nanobiosensorics Laboratory, Research Centre for Energy Research, Institute for Technical Physics and Materials Science, Konkoly-Thege u 29-33, 1120 Budapest, Hungary

Nowadays single-cell techniques are becoming valuable tools in the field of biology and biophysics. By investigating at a cellular level, we can better understand the cellular heterogeneity and the possible subpopulations in a tissue. This knowledge can bring novel applications and solutions to health and medicine. Fluidic Force Microscopy (FluidFM) is similar to atomic force microscopy (AFM), but uses a hollow, microfabricated cantilevers connected to a liquid reservoir and pressure controller system [1]. Through its precise force control, the cantilever is ideal to approach individual cells gently and reproducibly. Depending on the applications, the end of the cantilever can be different. For cell adhesion measurements 2-8 μm circular openings are ideal [2], while for live cell sampling and injections [3][4], so-called nanosyringes are used. These cantilevers have pyramidal tips with a 600 nm opening at their side. The cantilevers used for cell adhesion measurements can have different spring constants, which also determine some applications of the device. The short lecture will introduce a robotic version of the technique by presenting experimental results on the adhesive and mechanical properties of both cancerous and healthy cell types.

Acknowledgements

„Supported by the ÚNKP-22-2 New National Excellence Program of the Ministry for Culture and Innovation from the source of the National Research, Development and Innovation Fund.”

References

[1]: Li, M., Liu, L. & Zambelli, T. FluidFM for single-cell biophysics. Nano Res. 15, 773–786 (2022). https://doi.org/10.1007/s12274-021-3573-y

[2]: Sztilkovics, M., Gerecsei, T., Peter, B. et al. Single-cell adhesion force kinetics of cell populations from combined label-free optical biosensor and robotic fluidic force microscopy. Sci Rep 10, 61 (2020). https://doi.org/10.1038/s41598-019-56898-7

[3]: Chen, W., Guillaume-Gentil, O., Rainer, P.Y. et al. Live-seq enables temporal transcriptomic recording of single cells. Nature 608, 733–740 (2022). https://doi.org/10.1038/s41586-022-05046-9

[4]: Robert Horvath,Single-cell temporal transcriptomics from tiny cytoplasmic biopsies, Cell Reports Methods,Volume 2, Issue 10, 2022, 100319, ISSN 2667-2375, https://doi.org/10.1016/j.crmeth.2022.100319.