Detection of surface brush on biological cells <i>in vitro</i> with atomic force microscopy

Sokolov, Igor; Iyer, Swaminathan Padmanabhan; Subba-Rao, Venkatesh; Gaikwad, Ravi; Woodworth, Craig D.

doi:10.1063/1.2757104

Cited by 99 publications

(140 citation statements)

References 16 publications

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“…This means that the Young's modulus of the cell is not constant over the whole range of indentation. The same observations were made by Kasas et al and by Sokolov et al while indenting COS cells [32] and human cervical epithelial cells [61], respectively. These authors considered the cell as a mechanically multilayered structure in which the first layer (the most superficial) represents the actin cytoskeleton [32] or molecular brushes (microvilli, microridges, glycocalyx) [61], and the second layer represents the intermediate filament and microtubule network or "bulky cytosol".…”

Section: Hertz Modelsupporting

confidence: 81%

“…The same observations were made by Kasas et al and by Sokolov et al while indenting COS cells [32] and human cervical epithelial cells [61], respectively. These authors considered the cell as a mechanically multilayered structure in which the first layer (the most superficial) represents the actin cytoskeleton [32] or molecular brushes (microvilli, microridges, glycocalyx) [61], and the second layer represents the intermediate filament and microtubule network or "bulky cytosol". It is likely that in our experiments also that the 'membrane zone' (membrane ruffles, cortical actin cytoskeleton, lipid bilayer, membrane proteins) accounts for the first linear slope (E 1 ) and that the second slope (E 2 ) is caused by the elasticity of the bulky cytosol (microtubule network, intermediate filament network, cell organelles).…”

Section: Hertz Modelsupporting

confidence: 81%

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Elasticity measurement of living cells with an atomic force microscope: data acquisition and processing

Carl

Schillers

2008

Pflugers Arch - Eur J Physiol

208

176

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Elasticity of living cells is a parameter of increasing importance in cellular physiology, and the atomic force microscope is a suitable instrument to quantitatively measure it. The principle of an elasticity measurement is to physically indent a cell with a probe, to measure the applied force, and to process this force-indentation data using an appropriate model. It is crucial to know what extent the geometry of the indenting probe influences the result. Therefore, we indented living Chinese hamster ovary cells at 37 degrees C with sharp tips and colloidal probes (spherical particle tips) of different sizes and materials. We furthermore developed an implementation of the Hertz model, which simplifies the data processing. Our results show (a) that the size of the colloidal probe does not influence the result over a wide range (radii 0.5-26 microm) and (b) indenting cells with sharp tips results in higher Young's moduli ( approximately 1,300 Pa) than using colloidal probes ( approximately 400 Pa).

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Section: Hertz Modelsupporting

confidence: 81%

Section: Hertz Modelsupporting

confidence: 81%

Elasticity measurement of living cells with an atomic force microscope: data acquisition and processing

Carl

Schillers

2008

Pflugers Arch - Eur J Physiol

208

176

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“…As was shown in [52], it is critically important to take into account the pericellular coat for proper determination of biophysical properties of cells in the AFM indentation experiments. While the presence of the pericellular layer is well known, its nontrivial contribution to cell mechanics has only recently been confirmed [6, 47, 48, 53].…”

Section: Methodsmentioning

confidence: 99%

AFM study shows prominent physical changes in elasticity and pericellular layer in human acute leukemic cells due to inadequate cell–cell communication

et al. 2016

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Biomechanical properties of single cells in-vitro or ex-vivo and its pericellular interface have recently attracted a lot of attention as a potential biophysical (and possibly prognostic) marker of various diseases and cell abnormalities. At the same time, the influence of the cell environment on the biomechanical properties of cells is not well studied. Here we use atomic force microscopy (AFM) to demonstrate that cell-cell communication can have a profound effect on both cell elasticity and its pericellular coat. A human pre-B p190BCR/ABL acute lymphoblastic leukemia cell line (ALL3) was used in this study. Assuming that cell-cell communication is inversely proportional to the distance between cells, we study ALL3 cells in-vitro growing at different cell densities. ALL3 cells demonstrate a clear density dependent behavior. These cells grow very well if started at a relatively high cell density (HD, >2×105 cells/ml)) and are poised to grow at low cell density (LD, <1×104 cells/ml). Here we observe ~6x increase in the elastic (Young's) modulus of the cell body and ~3.6x decrease in the pericellular brush length of LD cells compared to HD ALL3 cells. The difference observed in the elastic modulus is much larger than typically reported for pathologically transformed cells. Thus, cell-cell communication must be taken into account when studying biomechanics of cells, in particular, correlating cell phenotype and its biophysical properties.

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“…Various membrane protrusions, which can be seen in optical confocal microscopy (see, e.g., 20 ) can be detected with AFM 33 . It has recently been found that the pericellular brush interferes with indentation measurements of elastic properties of the cell body, and a new model must be used 34 which separates contribution of the pericellular brush layer and deformation of the cell body in the AFM indentation experiments. Interestingly, cancer cells may look artificially softer if the cellular brush is not taken into account as was shown for the case of human cervical epithelial cells 20 .…”

Section: Introductionmentioning

confidence: 99%

Biophysical differences between chronic myelogenous leukemic quiescent and proliferating stem/progenitor cells

Guz

Patel

Dokukin

et al. 2016

Nanomedicine: Nanotechnology, Biology and Medicine

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The treatment of chronic myeloid leukemia (CML), a clonal myeloproliferative disorder has improved recently, but most patients have not yet been cured. Some patients develop resistance to the available tyrosine kinase treatments. Persistence of residual quiescent CML stem cells (LSC) that later resume proliferation is another common cause of recurrence or relapse of CML. Eradication of quiescent LSCs is a promising approach to prevent recurrence of CML. Here we report on new biophysical differences between quiescent and proliferating CD34+ LSCs, and speculate how this information could be of use to eradicate quiescent LSCs. Using AFM measurements on cells collected from four untreated CML patients, substantial differences are observed between quiescent and proliferating cells in the elastic modulus, pericellular brush length and its grafting density at the single cell level. The higher pericellular brush densities of quiescent LSCs are common for all samples. The significance of these observations is discussed.

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Detection of surface brush on biological cells in vitro with atomic force microscopy

Cited by 99 publications

References 16 publications

Elasticity measurement of living cells with an atomic force microscope: data acquisition and processing

Elasticity measurement of living cells with an atomic force microscope: data acquisition and processing

AFM study shows prominent physical changes in elasticity and pericellular layer in human acute leukemic cells due to inadequate cell–cell communication

Biophysical differences between chronic myelogenous leukemic quiescent and proliferating stem/progenitor cells

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