Fast automated processing of AFM PeakForce curves to evaluate spatially resolved Young modulus and stiffness of turgescent cells

Offroy, Marc; Razafitianamaharavo, Angélina; Beaussart, Audrey; Pagnout, Christophe; Duval, Jérôme F. L.

doi:10.1039/d0ra00669f

Cited by 23 publications

(25 citation statements)

References 73 publications

(132 reference statements)

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Section: Resultsmentioning

confidence: 99%

“…Spatial-distributions of the cell Young modulus ( E in Pa) and cell stiffness ( k cell in Nm −1 ), indicative of the cell Turgor pressure 46 , 50 , were evaluated for (un)exposed JW3606 (hep+) and JW3596 (hep−) (Figs. 6a, b (i),(ii) and 7a, b (i),(ii), respectively) from theoretical analysis 50 of 65,536 approach force curves collected by atomic force spectroscopy operated in PeakForce Tapping mode on 500 × 500 nm 2 single-cell surface areas (see details in “Methods”). Below, we further introduce δ defined by the value of the indentation (in nm) which marks the transition between the non-linear elastic deformation of the cell envelope and the linear compliance domain in the force versus indentation curve measured at a given location (pixel) of the cell surface 50 .…”

Section: Resultsmentioning

confidence: 99%

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Osmotic stress and vesiculation as key mechanisms controlling bacterial sensitivity and resistance to TiO2 nanoparticles

et al. 2021

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Toxicity mechanisms of metal oxide nanoparticles towards bacteria and underlying roles of membrane composition are still debated. Herein, the response of lipopolysaccharide-truncated Escherichia coli K12 mutants to TiO2 nanoparticles (TiO2NPs, exposure in dark) is addressed at the molecular, single cell, and population levels by transcriptomics, fluorescence assays, cell nanomechanics and electrohydrodynamics. We show that outer core-free lipopolysaccharides featuring intact inner core increase cell sensitivity to TiO2NPs. TiO2NPs operate as membrane strippers, which induce osmotic stress, inactivate cell osmoregulation and initiate lipid peroxidation, which ultimately leads to genesis of membrane vesicles. In itself, truncation of lipopolysaccharide inner core triggers membrane permeabilization/depolarization, lipid peroxidation and hypervesiculation. In turn, it favors the regulation of TiO2NP-mediated changes in cell Turgor stress and leads to efficient vesicle-facilitated release of damaged membrane components. Remarkably, vesicles further act as electrostatic baits for TiO2NPs, thereby mitigating TiO2NPs toxicity. Altogether, we highlight antagonistic lipopolysaccharide-dependent bacterial responses to nanoparticles and we show that the destabilized membrane can generate unexpected resistance phenotype.

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Section: Resultsmentioning

confidence: 99%

Section: Resultsmentioning

confidence: 99%

Osmotic stress and vesiculation as key mechanisms controlling bacterial sensitivity and resistance to TiO2 nanoparticles

et al. 2021

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“…In addition, self‐written software can be used to detect and calculate adhesion forces and energies [ 81 ] as well as elastic moduli and stiffness. [ 82 ]…”

Section: Resultsmentioning

confidence: 99%

Fundamentals and Applications of FluidFM Technology in Single‐Cell Studies

Saha

Duanis‐Assaf

Reches

2020

Adv Materials Inter

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various microfluidic devices have been routinely employed. However, in these approaches, there is either a high chance of damaging the cells while detaching the adherent cells from the surfaces, or isolation of individual cells from the adherent cell culture is not at all possible. Table 1 lists the drawbacks and challenges associated with these cell sorting techniques. Recently, atomic force microscope (AFM) has emerged as a significant tool to study microorganisms and mammalian cells in real time. This is due to its compatibility with the physiological conditions of the cells. [3] AFM allows the bioimaging of cells with various degrees of resolution, from whole cells [4] down to individual molecules. [5] Using force-curve based imaging, Alsteens et al. revealed single bacteriophages extruding from the surface of bacteria, [6] whereas our group was able to image nanostructures termed microvilli on the surface of T cells. [7] With the introduction of high-speed AFMs, Yamashita et al. were able to follow the diffusion of single molecules on the surface of living bacterial cells in real time. [8] Valuable biophysical and structural information can be achieved down to a single-cell level by means of high-resolution imaging and force spectroscopy. [9] In force spectroscopy experiments using AFM, a cell is directly attached to the cantilever apex with a biocompatible glue (e.g., polydopamine (PDA)). However, since the cell is irreversibly bound to the AFM probe, each functionalized cantilever can be utilized with only one cell for manipulation. It is, therefore, challenging to perform serial and rapid measurement of a reasonable number of cells; this makes it a time-consuming and elaborate process to address a statistical distribution. For injecting biomolecules such as dyes, enzymes or nanoparticles into single living cells, various approaches such as nano-fountain probes, [10] nanoneedles, [11] and carbon nanotubes [12] have been used. However, these approaches lack precise handling for local dispensing of fluid and are limited to the handling of very small volumes (on the order of attoliters (aLs) up to 50 femtoliters (fLs)). Moreover, using these approaches, a relatively long period of time is required to perform intracellular injection. With the advancement of technologies, it has become relatively easier to address these problems in a more controlled manner. Conventional AFM has been integrated with microfluidics to overcome these limitations. [13] This bioanalytical tool is known as a fluidic force microscope or FluidFM, which was This review describes the potential of FluidFM technology and its implementation in studying the interface between a single cell (prokaryote or eukaryote) and a surface or a surrounding area. A combination of microfluidics with conventional atomic force microscope (AFM) makes this platform efficient to address challenges associated with various biomolecular systems and biophysical activities down to single-cell levels. Upon regulating the pressure through a microchanneled cantilever v...

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“…E = σ/ε, the ratio of stress to strain. [7,17,18,38,39,43,48,69,70,[105][106][107]123,125,126,133,134,[139][140][141][142][143]149,152,155] Shear modulus (G)…”

Section: Terminology Explanation Common Formula or Law Referencementioning

confidence: 99%

“…Cell mechanics involves the study of the deformation of cell membrane and cytoskeleton, rigidity, viscoelasticity, adhesion force, and other mechanical properties of cells under mechanical loading, as well as the influence of mechanical factors on cell growth, development, maturation, proliferation, aging, and death. For disease research, the origin of many diseases can be considered from the mechanical properties of cells [4,5], such as the change of viscoelasticity of cells [6] and the change of Young's modulus [7], which are related to cancer metastasis and malignant transformation.…”

Section: Introductionmentioning

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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Fast automated processing of AFM PeakForce curves to evaluate spatially resolved Young modulus and stiffness of turgescent cells

Abstract: A numerical method is proposed for the modeling of AFM PeakForce curves and the automated extraction of relevant spatially-resolved nanomechanical properties of turgescent cells.

Cited by 23 publications

References 73 publications

Osmotic stress and vesiculation as key mechanisms controlling bacterial sensitivity and resistance to TiO2 nanoparticles

Osmotic stress and vesiculation as key mechanisms controlling bacterial sensitivity and resistance to TiO2 nanoparticles

Fundamentals and Applications of FluidFM Technology in Single‐Cell Studies

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

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