Adhesion force measurements on functionalized microbeads: An in-depth comparison of computer controlled micropipette and fluidic force microscopy

Gerecsei, Tamás; Erdődi, István; Péter, Beatrix; Hős, Csaba; Kurunczi, Sándor; Derényi, Imre; Szabó, Bálint; Horváth, Róbert

doi:10.1016/j.jcis.2019.07.102

Cited by 26 publications

(23 citation statements)

References 41 publications

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“…**when calibrated using FluidFM BOT 41 . ***This value is based on numerical simulations on the lifting force of microbeads 60 , and on direct comparison with FluidFM BOT force data 53 .…”

Section: Introductionmentioning

confidence: 99%

“…Both of them are very low throughput methods. Note, using a modified AFM applying vacuum on cells with a fluidic micro-channel (Fluidic Force Microscopy, FluidFM), the throughput is increased by a factor of 10 compared to conventional AFM 36 , 50 – 53 (Table 1 ). The optical signal of a high spatial resolution RWG (Single-Cell RWG) was recently calibrated by our group to adhesion force and energy values using a robotic fluidic force microscopy setup (FluidFM BOT), making possible to resolve the kinetics of single-cell adhesion forces and energy of large cell populations 41 .…”

Section: Introductionmentioning

confidence: 99%

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Single-cell adhesion strength and contact density drops in the M phase of cancer cells

Ungai-Salánki

Haty

Gerecsei

et al. 2021

Sci Rep

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The high throughput, cost effective and sensitive quantification of cell adhesion strength at the single-cell level is still a challenging task. The adhesion force between tissue cells and their environment is crucial in all multicellular organisms. Integrins transmit force between the intracellular cytoskeleton and the extracellular matrix. This force is not only a mechanical interaction but a way of signal transduction as well. For instance, adhesion-dependent cells switch to an apoptotic mode in the lack of adhesion forces. Adhesion of tumor cells is a potential therapeutic target, as it is actively modulated during tissue invasion and cell release to the bloodstream resulting in metastasis. We investigated the integrin-mediated adhesion between cancer cells and their RGD (Arg-Gly-Asp) motif displaying biomimetic substratum using the HeLa cell line transfected by the Fucci fluorescent cell cycle reporter construct. We employed a computer-controlled micropipette and a high spatial resolution label-free resonant waveguide grating-based optical sensor calibrated to adhesion force and energy at the single-cell level. We found that the overall adhesion strength of single cancer cells is approximately constant in all phases except the mitotic (M) phase with a significantly lower adhesion. Single-cell evanescent field based biosensor measurements revealed that at the mitotic phase the cell material mass per unit area inside the cell-substratum contact zone is significantly less, too. Importantly, the weaker mitotic adhesion is not simply a direct consequence of the measured smaller contact area. Our results highlight these differences in the mitotic reticular adhesions and confirm that cell adhesion is a promising target of selective cancer drugs as the vast majority of normal, differentiated tissue cells do not enter the M phase and do not divide.

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“…**when calibrated using FluidFM BOT 41 . ***This value is based on numerical simulations on the lifting force of microbeads 60 , and on direct comparison with FluidFM BOT force data 53 .…”

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Single-cell adhesion strength and contact density drops in the M phase of cancer cells

Ungai-Salánki

Haty

Gerecsei

et al. 2021

Sci Rep

Self Cite

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“…In different kinds of cells, the measurement of mechanical properties is widely used [65]. Gerecsei [66] used a computer to control micropipettes so as to achieve high-throughput measurement of the surface adhesion characteristics of particles. Hirano [67] established a feedback control system by combining scanning electron microscopy and micropipette technology.…”

Section: Micropipette Aspiration Techniquementioning

confidence: 99%

Recent Advances on the Model, Measurement Technique, and Application of Single Cell Mechanics

Huang

Dai

Shen

et al. 2020

IJMS

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Since the cell was discovered by humans, it has been an important research subject for researchers. The mechanical response of cells to external stimuli and the biomechanical response inside cells are of great significance for maintaining the life activities of cells. These biomechanical behaviors have wide applications in the fields of disease research and micromanipulation. In order to study the mechanical behavior of single cells, various cell mechanics models have been proposed. In addition, the measurement technologies of single cells have been greatly developed. These models, combined with experimental techniques, can effectively explain the biomechanical behavior and reaction mechanism of cells. In this review, we first introduce the basic concept and biomechanical background of cells, then summarize the research progress of internal force models and experimental techniques in the field of cell mechanics and discuss the latest mechanical models and experimental methods. We summarize the application directions of cell mechanics and put forward the future perspectives of a cell mechanics model.

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“…There are several methods for characterizing cell adhesion on such functionalized surfaces [15]. An emerging technique operating on the single-cell level is the computercontrolled micropipette [16,17], which is based on a glass micropipette connected to a pressure controller system. Adhered cells on a substrate are automatically directed under the micropipette, which attempts to aspire them using a negative pressure-induced flow.…”

Section: Introductionmentioning

confidence: 99%

“…The micropipette-based method also entails an important versatility in the types of samples that can be examined. This includes various cell types [16], but also other non-biological samples such as functionalized microbeads [17] as well. The latter can be used as a colloidal force spectroscopy tool to investigate the characteristics of immobilized receptor-ligand pairs.…”

Section: Introductionmentioning

confidence: 99%

Dissociation Constant of Integrin-RGD Binding in Live Cells from Automated Micropipette and Label-Free Optical Data

et al. 2021

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The binding of integrin proteins to peptide sequences such as arginine-glycine-aspartic acid (RGD) is a crucial step in the adhesion process of mammalian cells. While these bonds can be examined between purified proteins and their ligands, live-cell assays are better suited to gain biologically relevant information. Here we apply a computer-controlled micropipette (CCMP) to measure the dissociation constant (Kd) of integrin-RGD-binding. Surface coatings with varying RGD densities were prepared, and the detachment of single cells from these surfaces was measured by applying a local flow inducing hydrodynamic lifting force on the targeted cells in discrete steps. The average behavior of the populations was then fit according to the chemical law of mass action. To verify the resulting value of Kd2d = (4503 ± 1673) 1/µm2, a resonant waveguide grating based biosensor was used, characterizing and fitting the adhesion kinetics of the cell populations. Both methods yielded a Kd within the same range. Furthermore, an analysis of subpopulations was presented, confirming the ability of CCMP to characterize cell adhesion both on single cell and whole population levels. The introduced methodologies offer convenient and automated routes to quantify the adhesivity of living cells before their further processing.

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Adhesion force measurements on functionalized microbeads: An in-depth comparison of computer controlled micropipette and fluidic force microscopy

Cited by 26 publications

References 41 publications

Single-cell adhesion strength and contact density drops in the M phase of cancer cells

Single-cell adhesion strength and contact density drops in the M phase of cancer cells

Recent Advances on the Model, Measurement Technique, and Application of Single Cell Mechanics

Dissociation Constant of Integrin-RGD Binding in Live Cells from Automated Micropipette and Label-Free Optical Data

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