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Reference-Free Traction Force Microscopy Platform Fabricated via Two-Photon Laser Scanning Lithography Enables Facile Measurement of Cell-Generated Forces.

ABSTRACT: Cells sense and respond to the physical nature of their microenvironment by mechanically probing their surroundings via cytoskeletal contractions. The material response to these stresses can be measured via traction force microscopy (TFM). Traditional TFM platforms present several limitations including variable spatial resolution, difficulty in attaining the full three-dimensional (3D) deformation/stress profile, and the requirement to remove or relax the cells being measured to determine the zero-stress state. To overcome these limitations, we developed a two-photon, photochemical coupling approach to fabricate a new TFM platform that provides high-resolution control over the 3D placement of fluorescent fiducial markers for facile measurement of cell-generated shear and normal components of traction forces. The highly controlled placement of the 3D marker array provides a built-in, zero stress state eliminating the need to perturb the cells being measured while also providing increased throughput. Using this platform, we discovered that the magnitude of cell-generated shear and normal force components are linked both spatially and temporally. The facile nature and increased throughput of measuring cell-generated forces afforded by this new platform will be useful to the mechanotransduction community and others.

SUBMITTER: Banda OA

PROVIDER: S-EPMC8725169 | biostudies-literature |

REPOSITORIES: biostudies-literature

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Project description:Embedding liquid metals (LMs) into an elastomer is emerging as a promising strategy for stretchable conductors. Existing manufacturing techniques are struggling between spatial resolution and process complexity and are limited to chemically resistant substrates. Here, we report on a hybrid process combining stencil lithography and centrifugal force-assisted patterning of liquid metal for the development of LM-based stretchable conductors. The selective wetting behavior of oxide-removed eutectic gallium-indium (EGaIn) on metal patterns defined by stencil lithography enables micrometer scale LM patterns on an elastomeric substrate. Stencil lithography allows for defining metal regions without harsh chemical treatments, making it suitable for a wide range of substrates. Microscale LM patterns are achieved by efficiently removing the excess material by the centrifugal forces experienced from spinning the substrate. The proposed approach allows for the creation of LM patterns with a line width as small as 2 μm on a stretchable poly(dimethylsiloxane) (PDMS) substrate. The electrical measurement results show that the fabricated EGaIn devices can endure 40% mechanical strain over several thousands of cycles. Furthermore, a stencil design using microbridges is proposed to address the mechanical stability issue in stencil lithography. An EGaIn conductor with a serpentine structure and an interdigitated capacitor are fabricated and characterized. The results demonstrate that the patterned serpentine conductors retain their functionality with applied mechanical strain up to 80%. The performance of the interdigitated capacitors upon applied strain is in good agreement with the theoretical estimation. Finally, we demonstrate our approach also on poly(octamethylene maleate (anhydride) citrate) (POMaC) substrates to broaden the use of the proposed method to not only flexible and stretchable but also biodegradable substrates, opening a way for in vivo transient microsystem engineering. The work presented here provides a versatile and reliable approach for manufacturing stretchable conductors.

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Reference-Free Traction Force Microscopy Platform Fabricated via Two-Photon Laser Scanning Lithography Enables Facile Measurement of Cell-Generated Forces.

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