AFM‐Based Single‐Cell Force Spectroscopy

Franz, Clemens M.; Taubenberger, Anna

doi:10.1002/9783527649808.ch12

Cited by 5 publications

(10 citation statements)

References 63 publications

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“…Single‐cell force spectroscopy is gaining increasing popularity as technique to quantitate single‐cell adhesion forces (Helenius et al ., ; Franz and Taubenberger, ; Friedrichs et al ., ) but has so far been mainly used on homogenous substrates. However, integrating two or more different adhesive surfaces on the same substrate offers the opportunity to directly compare the adhesion strength of a single cell to different substrates (Dao et al ., ).…”

Section: Resultsmentioning

confidence: 99%

“…However, cell population behavior is often heterogeneous, requiring single‐cell adhesion assays to better describe the full spectrum of adhesion properties of a cell population. Over the last years, atomic force microscopy (AFM)‐based single‐cell force spectroscopy (SCFS) has evolved as a sensitive technique for quantitating single‐cell adhesion (Helenius et al ., ; Franz and Taubenberger, ). AFM‐based SCFS provides a versatile force range (~10 pN–100 nN), and the precise positioning system of the AFM allows excellent control over the contact conditions (interaction force and time).…”

Section: Introductionmentioning

confidence: 99%

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Investigating differential cell‐matrix adhesion by directly comparative single‐cell force spectroscopy

Dao

Gonnermann

Franz

2013

J of Molecular Recognition

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Tissue-embedded cells are often exposed to a complex mixture of extracellular matrix (ECM) molecules, to which they bind with different cell adhesion receptors and affinities. Differential cell adhesion to ECM components is believed to regulate many aspects of tissue function, such as the sorting of specific cell types into different tissue compartments or ECM niches. In turn, aberrant switches in cell adhesion preferences may contribute to cell misplacement, tissue invasion, and metastasis. Methods to determine differential adhesion profiles of single cells are therefore desirable, but established bulk assays usually only test cell population adhesion to a single type of ECM molecule. We have recently demonstrated that atomic force microscopy-based single-cell force spectroscopy (SCFS), performed on bifunctional, microstructured adhesion substrates, provides a useful tool for accurately quantitating differential matrix adhesion of single Chinese hamster ovary cells to laminin and collagen I. Here, we have extended this approach to include additional ECM substrates, such as bifunctional collagen I/collagen IV surfaces, as well as adhesion-passivated control surfaces. We investigate differential single cell adhesion to these substrates and analyze in detail suitable experimental conditions for comparative SCFS, including optimal cell-substrate contact times and the impact of force cycle repetitions on single cell adhesion force statistics. Insight gained through these experiments may help in adapting this technique to other ECM molecules and cell systems, making directly comparative SCFS a versatile tool for comparing receptor-mediated cell adhesion to different matrix molecules in a wide range of biological contexts.

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Section: Resultsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Investigating differential cell‐matrix adhesion by directly comparative single‐cell force spectroscopy

Dao

Gonnermann

Franz

2013

J of Molecular Recognition

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“…After varying contact times, the cell is retracted and a force‐distance curve is recorded. From the obtained curves, the maximum adhesion force ( F max ) and the detachment work ( W ) can be calculated . The maximum adhesion force F max , occurring at the moment of maximum AFM cantilever deflection, usually provides a suitable measure for the strength of cell adhesion on homogenous surfaces, while the deadhesion work W may also take in the contribution for individual adhesion contacts in mediating cell adhesion on structured cell adhesion substrates …”

Section: Resultsmentioning

confidence: 99%

Nanoscale topographic changes on sterilized glass surfaces affect cell adhesion and spreading

Wittenburg

Lauer

Oswald

et al. 2013

J Biomedical Materials Res

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Producing sterile glass surfaces is of great importance for a wide range of laboratory and medical applications, including in vitro cell culture and tissue engineering. However, sterilization may change the surface properties of glass and thereby affect its use for medical applications, for instance as a substrate for culturing cells. To investigate potential effects of sterilization on glass surface topography, borosilicate glass coverslips were left untreated or subjected to several common sterilization procedures, including low-temperature plasma gas, gamma irradiation and steam. Imaging by atomic force microscopy demonstrated that the surface of untreated borosilicate coverslips features a complex landscape of microislands ranging from 1000 to 3000 nm in diameter and 1 to 3 nm in height. Steam treatment completely removes these microislands, producing a nanosmooth glass surface. In contrast, plasma treatment partially degrades the microisland structure, while gamma irradiation has no effect on microisland topography. To test for possible effects of the nanotopographic structures on cell adhesion, human gingival fibroblasts were seeded on untreated or sterilized glass surfaces. Analyzing fibroblast adhesion 3, 6, and 24 h after cell seeding revealed significant differences in cell attachment and spreading depending on the sterilization method applied. Furthermore, single-cell force spectroscopy revealed a connection between the nanotopographic landscape of glass and the formation of cellular adhesion forces, indicating that fibroblasts generally adhere weakly to nanosmooth but strongly to nanorough glass surfaces. Nanotopographic changes induced by different sterilization methods may therefore need to be considered when preparing sterile glass surfaces for cell culture or biomedical applications.

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“…Cellular adhesion to surfaces (both host cells and invasive bacteria) controls many biological processes including inflammation and wound healing, infection, inter-cellular communication, tumor growth and metastasis, amongst others. 120,121) As its name suggests, single-cell force spectroscopy (SCFS) involves attachment of a living cell to an AFM cantilever, effectively turning the cell into a probe [Fig. 10(a)].…”

Section: Single-cell Force Spectroscopymentioning

confidence: 99%

Recent advances in micromechanical characterization of polymer, biomaterial, and cell surfaces with atomic force microscopy

Chyasnavichyus

Young

Tsukruk

2015

Jpn. J. Appl. Phys.

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Probing of micro- and nanoscale mechanical properties of soft materials with atomic force microscopy (AFM) gives essential information about the performance of the nanostructured polymer systems, natural nanocomposites, ultrathin coatings, and cell functioning. AFM provides efficient and is some cases the exclusive way to study these properties nondestructively in controlled environment. Precise force control in AFM methods allows its application to variety of soft materials and can be used to go beyond elastic properties and examine temperature and rate dependent materials response. In this review, we discuss experimental AFM methods currently used in the field of soft nanostructured composites and biomaterials. We discuss advantages and disadvantages of common AFM probing techniques, which allow for both qualitative and quantitative mappings of the elastic modulus of soft materials with nanosacle resolution. We also discuss several advanced techniques for more elaborate measurements of viscoelastic properties of soft materials and experiments on single cells.

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AFM‐Based Single‐Cell Force Spectroscopy

Cited by 5 publications

References 63 publications

Investigating differential cell‐matrix adhesion by directly comparative single‐cell force spectroscopy

Investigating differential cell‐matrix adhesion by directly comparative single‐cell force spectroscopy

Nanoscale topographic changes on sterilized glass surfaces affect cell adhesion and spreading

Recent advances in micromechanical characterization of polymer, biomaterial, and cell surfaces with atomic force microscopy

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