Összes szerző

Horváth Róbert

az alábbi absztraktok szerzői között szerepel:

Farkas Enikő
Controlling Live Cell Adhesion through Characterization of Biofunctionalized Surfaces using Label-Free Biosensors

Aug 30 - szerda

15:30 – 17:00

II. Poszterszekció

P41

Controlling Live Cell Adhesion through Characterization of Biofunctionalized Surfaces using Label-Free Biosensors

Eniko Farkas¹, Kinga Dóra Kovács^1,3, Beatrix Peter¹, Attila Bonyár², Sandor Kurunczi¹, Inna Szekacs¹, and Robert Horvath¹

¹ Nanobiosensorics Laboratory, Institute of Technical Physics and Materials Science, Centre for Energy Research, Budapest, Hungary

² Department of Electronics Technology, Faculty of Electrical Engineering and Informatics, Budapest University of Technology and Economics, Budapest, Hungary

Biomaterial coatings that possess cell-repellent or cell-adhesive properties have a significant interest in medical and biotechnological applications [1-4]. However, conventional approaches lack in-depth analysis and quantitative comparison of these coatings for regulating adhesion, particularly for bacterial cell adhesion. Label-free Optical Waveguide Lightmode Spectroscopy (OWLS) can offer a solution for the detailed analysis of biomaterial coatings. OWLS biosensors detect the optical properties of the adhesive surface using evanescent waves with a penetration depth of 100-150 nm [5-7]. This surface-sensitive technique enables a thorough evaluation of biomaterial coatings for regulating adhesion. Uniquely, OWLS enables the in situ measurement of both the coating process and subsequent cell adhesion.

The present study utilizes the OWLS method for in-depth characterization of biomaterial surfaces with regard to bacterial adhesion. Initially, adhesion blocking biomaterials, namely bovine serum albumin, I-block, PAcrAM-g-(PMOXA, NH2, Si), (PAcrAM-P), and PLL-g-PEG, with varying coating temperatures, were screened. PAcrAM-P exhibited the best blocking capability against bacterial concentrations up to 10⁷ cells/mL. Subsequently, different immobilization methods, such as Mix&Go (AnteoBind) films, protein A, avidin-biotin based surface chemistries, and simple physisorption, were employed to captureEscherichia coli specific antibodies. Bacterial cell adhesion was then tested on immobilized antibodies with various blocking agents. The OWLS analysis allowed for the determination of the parameters of the applied agents by considering the kinetic data of adhesion, the surface mass density, and the protein orientation. Based on the experimental results, surfaces were created and tested for controlling both bacterial and mammalian cell adhesion. [8]

Acknowledgment

This work was supported by the "Lendület" (HAS) research program, the National Research, Development and Innovation Office of Hungary ((ERC_HU, VEKOP 2.2.1-16, ELKH topic-fund, "Élvonal" KKP_19 and KH grants, PD 131543 and TKP2022-EGA-04 –INBIOM TKP Programs financed from the NRDI Fund). This work was also supported by 77 Elektronika Ltd. by their supplying of antibodies and reagents.

References

[1] Frutiger A, et. al. (2021) Chem Rev 121: 8095–8160.

[2] Rigo S, et. al. (2018) Adv Sci 5: 1700892.

[3] Castillo-Henríquez L, et. al. (2020) Sensors 20: 6926.

[4] D’Agata R, et. al. (2021) Polymers 13:1929.

[5] Vörös J, et. al. (2002) Biomaterials 23: 3699–3710.

[6] Tiefenthaler K, et. al. (1989) J Opt Soc Am. B 6: 209–220.

[7] Saftics A, et. al. (2021) Adv Colloid Interface Sci 294: 102431–102433.

[8] Farkas E, et. al. (2022) Biosensors 12: 56.

Kovács Boglárka
Flagellin: a convenient protein in biosensorics

Aug 30 - szerda

11:15 – 11:30

Bioszenzorika és bio-nanotechnológia

E27

Flagellin: a convenient protein in biosensorics

Boglárka Kovács¹, András Saftics¹, Inna Székács¹, Hajnalka Jankovics², Sandor Kurunczi¹, Ferenc Vonderviszt², Robert Horvath¹

¹Nanobiosensorics Laboratory, Centre for Energy Research, Institute of Technical Physics and Materials Science, Budapest, Hungary

²Bio-nanosystem Laboratory, Research Institute of Biomolecular and Chemical Engineering, University of Pannonia, Veszprém, Hungary

Flagellin is the main building block of bacterial flagellar filaments. Since the filaments are located outside of the cells, cell lysis is not required to purify flagellin. Flagellin consists of 4 domains: D0, D1, D2, and D3. The D0 domain contains amphipathic helical regions with hydrophobic amino acids on one side of the helix. This part of flagellin is disordered in solution, but can be used to anchor the protein on hydrophobic surfaces with the D3 domain pointing towards the solution [1]. The hypervariable D3 domain situated on the filament surface is a largely independent part of the flagellin that can be removed or replaced without disturbing filament formation.

During our work we in-depth characterized the coatings created from flagellin, and influenced the adsorption of the protein with Hofmeister salts [1]. We applied genetically modified high affinity Ni-binding variant as receptor, and demonstrated the unique sensitivity of grating-coupled interferometry [2].

The monolayer of wild-type flagellin mimics the surface of the bacterial flagellar filament, and we hypothesized that oriented flagellin layers have bacteria-repellent properties. To prove this, we studied the adhesion of bacterial E. coli and human cancer cells on oriented wild-type flagellin layers [3,4].

Through genetic modification, specific oligopeptide segments can be also inserted into the D3 domain of flagellin, which can induce cell adhesion through integrin receptors. We studied cancer cell adhesion on the genetically engineered protein layers with label-free optical biosensors [4]. Mammalian cells can recognize flagellin in solution through Toll-like receptors, and the protein can cause innate immune system response. We are studying the above biological mechanisms and its consequences in the adhesion of the flagelljn exposed cells. Our results prove, that flagellin can be used in many ways in creating capture layers in biosensors.

References

[1] Kovacs, B.; Saftics, A.; Biro, A.; Kurunczi, S.; Szalontai, B.; Kakasi, B.; Vonderviszt, F.; Der, A.; Horvath, R. J. Phys. Chem. C 2018, 122 (37), 21375–21386.

[2] Jankovics, H.; Kovacs, B.; Saftics, A.; Gerecsei, T.; Tóth, É.; Szekacs, I.; Vonderviszt, F.; Horvath, R. Sci. Rep. 2020, 1–11.

[3] Kovacs, B.; Patko, D.; Klein, A.; Kakasi, B.; Saftics, A.; Kurunczi, S.; Vonderviszt, F.; Horvath, R. Sensors Actuators B Chem. 2018, 257, 839–845.

[4] Kovacs, B.; Patko, D.; Szekacs, I.; Orgovan, N.; Kurunczi, S.; Sulyok, A.; Khanh, N. Q.; Toth, B.; Vonderviszt, F.; Horvath, R. Acta Biomater. 2016, No. 42, 66–76.

Kovács Dóra Kinga
Nanoinjection of fluorescent nanoparticles to single live cells by robotic fluidic force microscopy

Aug 29 - kedd

12:15 – 12:30

Modern biofizikai módszerek

E13

Nanoinjection of fluorescent nanoparticles to single live cells by robotic fluidic force microscopy

Kinga Dóra Kovács^1,*, Tamás Visnovitz^2,3,*, Tamás Gerecsei¹, Beatrix Peter¹, Sándor Kurunczi¹, Anna Koncz², Krisztina Németh², Dorina Lenzinger², Krisztina V. Vukman², Péter Lőrincz⁴, Inna Székács¹, Edit I. Buzás^2,5,6**, Robert Horvath^1,**

¹ Nanobiosensorics Laboratory, Centre of Energy Research, ELKH, Budapest, Hungary

² Department of Genetics, Cell- and Immunobiology, Semmelweis University, Budapest, Hungary

³ Department of Plant Physiology and Molecular Plant Biology, ELTE Eötvös Loránd University, Budapest, Hungary

⁴ Department of Anatomy, Cell and Developmental Biology, ELTE Eötvös Loránd University, Budapest, Hungary

⁵ HCEMM-SU Extracellular Vesicle Research Group, Budapest, Hungary

⁶ ELKH-SE Translational Extracellular Vesicle Research Group, Budapest, Hungary

*,** equal contributions / **corresponding authors

Direct injection of fluorescent nanoparticles into the cytoplasm of living cells can provide new insights into the intracellular fate of various different fluorescently labelled biologically active particles. Here we used fluorescent nanoparticles to prove the feasibility of nanoinjection into single live HeLa cells by using robotic fluidic force microscopy (FluidFM). This injection platform offers the advantage of high cell selectivity and efficiency. We confirmed the successful injection of both GFP encoding plasmids and GFP tagged fluorescent nanoparticles to the cells by confocal microscopy. We were able track the nanoparticles in the living cells for 20 hours. The injected nanoparticles were initially localized in concentrated spot-like regions within the cytoplasm. Later, they were transported towards the periphery of the cells. Based on our proof-of-principle data, the FluidFM platform is suitable for targeting single living cells by fluorescently labelled biologically active particles and may lead to information about the intracellular cargo delivery at a single-cell level.

Madas Balázs
Metastatic potential of HeLa-cells does not increase directly after radiation exposure

Aug 29 - kedd

14:20 – 14:40

Orvosi biofizika és sugárbiológia

E15

Metastatic potential of HeLa-cells does not increase directly after radiation exposure

Balázs Madas¹, Kinga Kovács², Andrea Strádi³, Szabolcs Polgár⁴, Inna Székács⁵, and Róbert Horváth⁵

¹ Centre for Energy Research, Environmental Physics Department, Budapest, Hungary

² Centre for Energy Research, Nanobiosensorics Department, Budapest, Hungary

³ Centre for Energy Research, Space Research Department, Budapest, Hungary

While radiation therapy increases local tumor control, it remains controversial whether ionizing radiation increases the metastatic potential of cancer cells. One of the potential mechanisms of radiation-induced metastasis is the direct release of tumor cells into the circulation requiring the detachment of the cells. The objective of the present study was to directly measure how ionizing radiation affects the kinetics of cellular adhesion, especially its initial stage after cell attachment on a biomimetic surface.

For this purpose, an automatic irradiation facility with gamma-radiation from Cs-137 has been developed providing parallel irradiation opportunity of 96 wells of a biosensor microplate with different doses. The employed optical biosensor records the wavelength shift of reflected light from a nanostructured waveguide, being proportional to the cell adhesion strength. Absorbed doses were measured by thermoluminescent dosimeters (TLDs) in each well. As a model system, a cervical cancer cell line (HeLa) was studied.

Three different experimental setups have been used distinguished by the sequence of irradiation and cellular attachment to the surface. The wavelength shift as the function of time was measured for different absorbed doses. The maximum wavelength shift as the function of dose was also analyzed.

The results show that adhesion of HeLa cells is not affected by ionizing radiation in the first hours after irradiation. The result is independent of whether cells are exposed during the adhesion process, in suspension, or attached. This suggests that radiation therapy does not directly increase the metastatic potential of cancer cells by decreasing their adhesion.

The experimental setup can be used to quantify the effects of ionizing radiation on cell adhesion as the function of time at different absorbed doses. It has been shown that ionizing radiation does not affect the adhesion of HeLa cells in the first hours after exposure, while experiments with longer follow-up are required to see whether adhesion changes at later time points.

Magyaródi Beatrix
Role of glycocalyx in cancer cell adhesion: kinetics of interactions from label-free optical biosensor measurements

Aug 30 - szerda

15:30 – 17:00

II. Poszterszekció

P46

Role of glycocalyx in cancer cell adhesion: kinetics of interactions from label-free optical biosensor measurements

Beatrix Magyaródi¹, Boglárka Kovács¹, Inna Székács¹, Robert Horvath¹

¹ Nanobiosensorics Laboratory, Research Centre for Energy Research, Institute for Technical Physics and Materials Science, Konkoly-Thege u 29-33, 1120 Budapest, Hungary

The glycocalyx is a sugar rich layer covering the surface of the cells.[1] It is composed of glycoproteins and proteoglycans. The cellular glycocalyx plays an important, but not yet understood, role in cellular signaling and metabolism, its disorders generate pathological process.[2] Interestingly, the thickness of the glycocalyx layer of cancer cells is significantly larger compared to that of healthy cells. This fact further highlights the importance of glycocalyx in tumor progression and treatment. In an earlier work, a regulatory mechanism of cellular glycocalyx in cancer adhesion was revealed using label-free optical biosensor, fluorescent microscopy, and cell surface charge measurements. [3]

The primary goal of our work is to study the role of glycocalyx components in cellular adhesion by employing various types of digesting methods. In these initial measurements we use a label-free, high-throughput, resonant waveguide grating-based optical biosensor. The instrument is well suited for monitoring of cellular adhesion kinetics in real-time, even at the single-cell level.[4]

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] M. J. Paszek et al., “The cancer glycocalyx mechanically primes integrin-mediated growth and survival,” Nature, vol. 511, no. 7509, pp. 319–325, 2014, doi: 10.1038/nature13535.

[2] E. R. Cruz-Chu, A. Malafeev, T. Pajarskas, I. V. Pivkin, and P. Koumoutsakos, “Structure and response to flow of the glycocalyx layer,” Biophys. J., 2014, doi: 10.1016/j.bpj.2013.09.060.

[3] N. Kanyo 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., 2020, doi: 10.1038/s41598-020-80033-6.

[4] M. Sztilkovics et al., “Single-cell adhesion force kinetics of cell populations from combined label-free optical biosensor and robotic fluidic force microscopy,” Sci. Rep., 2020, doi: 10.1038/s41598-019-56898-7.

Novák Szabolcs
Investigation of antibody-mediated adhesion force of individual immune cells using computer-controlled micropipette and fluidic force microscopy

Aug 29 - kedd

15:30 – 17:00

I. Poszterszekció

P19

Investigation of antibody-mediated adhesion force of individual immune cells using computer-controlled micropipette and fluidic force microscopy

S. Novák¹, Z. Szittner¹, I. Sallai¹, I. Székács¹, R. Horvath¹

¹ Nanobiosensorics Laboratory, Centre of Energy Research, Eötvös Loránd Research Network, Budapest, Hungary

Cell adhesion is a fundamental process that plays a critical role in various biological phenomena, such as the immune response. To understand the mechanisms of cell adhesion and develop new therapies, it is necessary to measure the forces involved.Computer-controlled micropipettes (CCMP) and Fluidic Force Microscopy (FluidFM) techniques have become effective instruments for the accurate determination of cell adhesion in recent years.

A CCMP is a tool that can be used to probe single cell interactions with specific macromolecules and surfaces. The micropipette is mounted onto a micromanipulator on a normal inverted microscope and is controlled by a computer. The adhesion force of surface-attached cells can be accurately probed by repeating a pick-up process on the examined cells while increasing a vacuum applied through a pump system in the pipette positioned above the cell. Using this methodology, high number of cells adhered to specific macromolecules, treated surfaces can be measured one by one in a short period of time. Additionally, the probed single cells can be easily picked up and separated for further examinations by other techniques. This is a definite advantage of the CCPM.[1]

FluidFM, on the other hand, uses a hollow cantilever with a small opening at the tip that can be filled with liquid using a fluid reservoir that is attached to a pressure control system, allowing for precise fluid dispensing and manipulation at the nanoscale. This technology enables the precise measurement of adhesion forces between cells and their targets and makes it possible to record the entire cell detachment process quickly and accurately. [2]

The combination of these two technologies provides high accuracy and resolution in the measurement of cell adhesion forces and in addition to measuring adhesion forces. We used these techniques to study the adhesion strength of immune cells on different surfaces.

Acknowledgment

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

References

[1] R. Salánki et al., “Single Cell Adhesion Assay Using Computer Controlled Micropipette,” PLoS One, vol. 9, no. 10, pp. e111450-, Oct. 2014, [Online]. Available: https://doi.org/10.1371/journal.pone.0111450

[2] Á. G. Nagy, I. Székács, A. Bonyár, and R. Horvath, “Cell-substratum and cell-cell adhesion forces and single-cell mechanical properties in mono- and multilayer assemblies from robotic fluidic force microscopy,” Eur J Cell Biol, vol. 101, no. 4, Sep. 2022, doi: 10.1016/j.ejcb.2022.151273.

Péter Beatrix
Nanoparticle uptake of living cells with digested glycocalyx

Aug 29 - kedd

15:30 – 17:00

I. Poszterszekció

P21

Nanoparticle uptake of living cells with digested glycocalyx

Beatrix Peter^a, Nicolett Kanyo^a, Kinga Dora Kovacs^a,b, Viktor Kovács^a, Inna Szekacs^a, Béla Pécz^c, Kinga Molnár^d,Hideyuki Nakanishi^e, Istvan Lagzi^,,f,g, Robert Horvath^a

^a Nanobiosensorics laboratory, Institute of Technical Physics and Materials Science, Centre for Energy Research, Konkoly-Thege út 29-33, H-1121 Budapest, Hungary

^b Department of Biological Physics, Eötvös University, Budapest, Hungary

^c Thin Films Laboratory, Institute of Technical Physics and Materials Science, Centre for Energy Research, Konkoly-Thege út 29-33, H-1120 Budapest, Hungary

^d Department of Anatomy, Cell and Developmental Biology, ELTE, Eötvös Loránd University, Pázmány Péter stny. 1/C, Budapest, H-1117, Hungary

^e Department of Macromolecular Science and Engineering, Graduate School of Science and Technology, Kyoto Institute of Technology, Matsugasaki, Kyoto 606-8585, Japan

^f Department of Physics, Institute of Physics, Budapest University of Technology and Economics, Műegyetem rkp. 3, Budapest H-1111, Hungary

^g ELKH BME Condensed Matter Research Group, Műegyetem rkp. 3, Budapest H-1111, Hungary

In biomedical imaging and targeted drug delivery, functionalized nanoparticles are widely used due to their penetration into living cells. The glycocalyx is a surface sugar layer of the cells, which presumably plays an essential role in any uptake process. However, its exact function in nanoparticle uptake is still uncovered. We in situ monitored the penetration of positively charged gold nanoparticles into adhered cancer cells with or without preliminary glycocalyx digestion. During the experiments, the components of glycocalyx of HeLa cells were digested by chondroitinase ABC enzyme. The measurements were performed by applying a high-throughput label-free resonant waveguide grating biosensor. The positively charged gold nanoparticles were used with different sizes (S, M, L). Negatively charged citrate-capped tannic acid nanoparticles, and other types of glycocalyx digesting enzymes were also applied in control experiments. The biosensor data confirmed the cellular uptake of the functionalized nanoparticles with an active process, which was verified by transmission electron microscopy [1,2]. Based on the findings we conclude that the components of gylcocalyx control the uptake process in size- and charge-dependent manner, and the possible roles of various glycocalyx components were highlighted.

Acknowledgements

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

References

[1] B. Peter, N. Kanyo, K. D. Kovacs, V. Kovács, I. Szekacs, B. Pécz, K. Molnár, H. Nakanishi, I. Lagzi, R. Horvath. Glycocalyx components detune the cellular uptake of gold nanoparticles in a size- and charge-dependent manner. ACS Applied Bio Materials, 2023.

[2] B. Peter, I. Lagzi, S. Teraji, H. Nakanishi, L. Cervenak, D. Zámbó, A. Deák, K. Molnár, M. Truszka, I. Szekacs, R. Horvath. Interaction of positively charged gold nanoparticles with cancer cells monitored by an in situ label-free optical biosensor and transmission electron microscopy. ACS Applied Materials & Interfaces, 2018.

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.

Sallai Igor
Label-free immune cell analysis using optical biosensor

Aug 30 - szerda

15:30 – 17:00

II. Poszterszekció

P52

Label-free immune cell analysis using optical biosensor

I. Sallai¹, Z. Szittner¹, Sz. Novák¹, I. Székács¹ and R. Horvath¹

¹ Nanobiosensorics Laboratory, Center of Energy Research, Eötvös Loránd Research Network, Budapest, Hungary

Understanding of activation processes at the single-cell level in response to different stimuli is essential for the diagnosis of certain diseases.

External stimuli induced differences are reflected in the dynamic changes of cell biophysical parameters, such as cell motility, shape, spreading and adhesion properties [1]. Novel highly sensitive optical biosensors allow the monitoring of changes in these parameters in a label-free manner. The advantage of label-free detection is that cells can be examined in an intact/organism-specific manner without modification.

The above parameters can be studied using microplate-based, high-throughput systems [2]. Surface functionalisation provide the opportunity to create an organism-specific environment [3]. In classical measurements, such as microscopy, dye-conjugated antibody labelling is essential and can be combined with label-free data [4].

Our aim is to interpret the activation mechanisms of various cell types and their function triggered by different stimuli and compare the results with conventional testing methods.

Acknowledgment

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

References

[1] Z. Szittner, B. Péter, S. Kurunczi, I. Székács, and R. Horvath, “Functional blood cell analysis by label-free biosensors and single-cell technologies,” Advances in Colloid and Interface Science, vol. 308. Elsevier B.V., Oct. 01, 2022. doi: 10.1016/j.cis.2022.102727.

[2] M. Sztilkovics et al., “Single-cell adhesion force kinetics of cell populations from combined label free optical biosensor and robotic fluidic force microscopy,” Sci Rep, vol. 10, no. 1, Dec. 2020, doi: 10.1038/s41598-019-56898-7.

[3] N. Orgovan et al., “In-situ and label-free optical monitoring of the adhesion and spreading of primary monocytes isolated from human blood: Dependence on serum concentration levels,” Biosens Bioelectron, vol. 54, pp. 339–344, Apr. 2014, doi: 10.1016/j.bios.2013.10.076.

[4] S. Zheng, J. C. H. Lin, H. L. Kasdan, and Y. C. Tai, “Fluorescent labeling, sensing, and differentiation of leukocytes from undiluted whole blood samples,” Sens Actuators B Chem, vol. 132, no. 2, pp. 558–567, Jun. 2008, doi: 10.1016/j.snb.2007.11.031.

Szittner Zoltán
Label-free single-cell compatible biophysical methods in immune cell activation

Aug 30 - szerda

12:00 – 12:15

Bioszenzorika és bio-nanotechnológia

E30

Label-free single-cell compatible biophysical methods in immune cell activation

Z. Szittner, S. Novák, I. Sallai, I. Székács, R. Horvath

Nanobiosensorics Laboratory, Centre of Energy Research, ELKH, Budapest, Hungary

Recent advances in biophysical methods provide a novel approach to characterize immune cell activation. Here, we present an experimental platform for studying adhesion kinetics, adhesion force, and cell morphology at a single-cell level. These techniques enable the precise characterization of complex cellular mixtures and the testing of the effects of various compounds on immune cell activation(1). The adhesion force, measured by computer-controlled micropipette and fluidic force microscopy, serves as a proxy for cellular activation induced by various compounds(2). The resonant wavelength grating technique exploits the sensitivity of the surface-bound optical evanescent field to changes in the local refractive index, enabling the study of cell adhesion kinetics in single cells at subminute time resolutions(3). Moreover, digital holographic microscopy records the morphology of single cells during their activation and extracts multiple features, such as cell area, optical thickness, and motility, to characterize their activation state(4). Importantly, these biophysical methods enable the characterization of cellular processes in a label-free manner, reducing the complexity and material demand of each measurement and enabling the investigation of single cells in their native state. Comparing and evaluating these techniques carefully can enhance our understanding of immune cell activation and lead to the development of diagnostic approaches and novel therapies for immune system-related questions.

Acknowledgements

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. Szittner Z, Péter B, Kurunczi S, Székács I, Horvath R. Functional blood cell analysis by label-free biosensors and single-cell technologies. Advances in Colloid and Interface Science. 2022 Oct;308:102727.

2. Ungai-Salánki R, Peter B, Gerecsei T, Orgovan N, Horvath R, Szabó B. A practical review on the measurement tools for cellular adhesion force. Advances in Colloid and Interface Science. 2019 Jul 1;269:309–33.

3. Sztilkovics M, Gerecsei T, Peter B, Saftics A, Kurunczi S, Szekacs I, et al. Single-cell adhesion force kinetics of cell populations from combined label-free optical biosensor and robotic fluidic force microscopy. Sci Rep. 2020 Jan 9;10(1):61.

4. Nagy ÁG, Székács I, Bonyár A, Horvath R. Simple and automatic monitoring of cancer cell invasion into an epithelial monolayer using label-free holographic microscopy. Sci Rep. 2022 Jun 16;12(1):10111.