Dataset Information

Nanonet Force Microscopy for Measuring Cell Forces.

ABSTRACT: The influence of physical forces exerted by or felt by cells on cell shape, migration, and cytoskeleton arrangement is now widely acknowledged and hypothesized to occur due to modulation of cellular inside-out forces in response to changes in the external fibrous environment (outside-in). Our previous work using the non-electrospinning Spinneret-based Tunable Engineered Parameters' suspended fibers has revealed that cells are able to sense and respond to changes in fiber curvature and structural stiffness as evidenced by alterations to focal adhesion cluster lengths. Here, we present the development and application of a suspended nanonet platform for measuring C2C12 mouse myoblast forces attached to fibers of three diameters (250, 400, and 800 nm) representing a wide range of structural stiffness (3-50 nN/?m). The nanonet force microscopy platform measures cell adhesion forces in response to symmetric and asymmetric external perturbation in single and cyclic modes. We find that contractility-based, inside-out forces are evenly distributed at the edges of the cell, and that forces are dependent on fiber structural stiffness. Additionally, external perturbation in symmetric and asymmetric modes biases cell-fiber failure location without affecting the outside-in forces of cell-fiber adhesion. We then extend the platform to measure forces of (1) cell-cell junctions, (2) single cells undergoing cyclic perturbation in the presence of drugs, and (3) cancerous single-cells transitioning from a blebbing to a pseudopodial morphology.

SUBMITTER: Sheets K

PROVIDER: S-EPMC4945583 | biostudies-literature | 2016 Jul

REPOSITORIES: biostudies-literature

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Nanonet Force Microscopy for Measuring Cell Forces.

Sheets Kevin K Wang Ji J Zhao Wei W Kapania Rakesh R Nain Amrinder S AS

Biophysical journal 20160701 1

The influence of physical forces exerted by or felt by cells on cell shape, migration, and cytoskeleton arrangement is now widely acknowledged and hypothesized to occur due to modulation of cellular inside-out forces in response to changes in the external fibrous environment (outside-in). Our previous work using the non-electrospinning Spinneret-based Tunable Engineered Parameters' suspended fibers has revealed that cells are able to sense and respond to changes in fiber curvature and structural ...[more]

PMID: 27410747

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Project description:BACKGROUND: Lignin is a complex polymer which inhibits the enzymatic conversion of cellulose to glucose in lignocellulose biomass for biofuel production. Cellulase enzymes irreversibly bind to lignin, deactivating the enzyme and lowering the overall activity of the hydrolyzing reaction solution. Within this study, atomic force microscopy (AFM) is used to compare the adhesion forces between cellulase and lignin with the forces between cellulase and cellulose, and to study the moiety groups involved in binding of cellulase to lignin. RESULTS: Trichoderma reesei, ATCC 26921, a commercial cellulase system, was immobilized onto silicon wafers and used as a substrate to measure forces involved in cellulase non-productive binding to lignin. Attraction forces between cellulase and lignin, and between cellulase and cellulose were compared using kraft lignin- and hydroxypropyl cellulose-coated tips with the immobilized cellulase substrate. The measured adhesion forces between kraft lignin and cellulase were on average 45% higher than forces between hydroxypropyl cellulose and cellulase. Specialized AFM tips with hydrophobic, -OH, and -COOH chemical characteristics were used with immobilized cellulase to represent hydrophobic, H-bonding, and charge-charge interactions, respectively. Forces between hydrophobic tips and cellulase were on average 43% and 13% higher than forces between cellulase with tips exhibiting OH and COOH groups, respectively. A strong attractive force during the AFM tip approach to the immobilized cellulase was observed with the hydrophobic tip. CONCLUSIONS: This work shows that there is a greater overall attraction between kraft lignin and cellulase than between hydroxypropyl cellulose and cellulase, which may have implications during the enzymatic reaction process. Furthermore, hydrophobic interactions appear to be the dominating attraction force in cellulase binding to lignin, while a number of other interactions may establish the irreversible binding.

Dataset Information

Nanonet Force Microscopy for Measuring Cell Forces.

Publications

Nanonet Force Microscopy for Measuring Cell Forces.

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