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A multi-material platform for imaging of single cell-cell junctions under tensile load fabricated with two-photon polymerization

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Abstract

We previously reported a single-cell adhesion micro tensile tester (SCAμTT) fabricated from IP-S photoresin with two-photon polymerization (TPP) for investigating the mechanics of a single cell-cell junction under defined tensile loading. A major limitation of the platform is the autofluorescence of IP-S, the photoresin for TPP fabrication, which significantly increases background signal and makes fluorescent imaging of stretched cells difficult. In this study, we report the design and fabrication of a new SCAμTT platform that mitigates autofluorescence and demonstrate its capability in imaging a single cell pair as its mutual junction is stretched. By employing a two-material design using IP-S and IP-Visio, a photoresin with reduced autofluorescence, we show a significant reduction in autofluorescence of the platform. Further, by integrating apertures onto the substrate with a gold coating, the influence of autofluorescence on imaging is almost completely mitigated. With this new platform, we demonstrate the ability to image a pair of epithelial cells as they are stretched up to 250% strain, allowing us to observe junction rupture and F-actin retraction while simultaneously recording the accumulation of over 800 kPa of stress in the junction. The platform and methodology presented here can potentially enable detailed investigation of the mechanics of and mechanotransduction in cell-cell junctions and improve the design of other TPP platforms in mechanobiology applications.

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Acknowledgements

We thank Dr. Benjamin Richter from Nanoscribe GmbH for the helpful discussions.

Funding

We acknowledge the funding support from the NSF (Awards 1826135, 1936065, 2143997), the NIH National Institutes of General Medical Sciences P20GM113126 (Nebraska Center for Integrated Biomolecular Communication) and P30GM127200 (Nebraska Center for Nanomedicine), the Nebraska Collaborative Initiative and the Voelte-Keegan Bioengineering Support. Design and fabrication of the TPP structures were conducted at the Center for Nanophase Materials Sciences (CNMS) at ORNL, which is a DOE Office of Science User Facility. Manufacturing and characterization analysis were performed at the NanoEngineering Research Core Facility (NERCF). G.M. and J.R. are funded by the NSF Graduate Research Fellowship (Award 2034837).

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Authors and Affiliations

  1. Department of Mechanical and Materials Engineering, University of Nebraska-Lincoln, Lincoln, NE, 68588, USA

    Jordan Rosenbohm, Grayson Minnick, Bahareh Tajvidi Safa, Amir Monemian Esfahani, Xiaowei Jin, Haiwei Zhai & Ruiguo Yang

  2. Center for Nanophase Materials Sciences, Oak Ridge National Laboratory, Oak Ridge, TN, 37831-6054, USA

    Nickolay V. Lavrik

  3. Nebraska Center for Integrated Biomolecular Communication, University of Nebraska-Lincoln, Lincoln, NE, 68588, USA

    Ruiguo Yang

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  1. Jordan Rosenbohm
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  2. Grayson Minnick
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  3. Bahareh Tajvidi Safa
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  4. Amir Monemian Esfahani
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  5. Xiaowei Jin
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  6. Haiwei Zhai
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  7. Nickolay V. Lavrik
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Correspondence to Nickolay V. Lavrik or Ruiguo Yang.

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Rosenbohm, J., Minnick, G., Safa, B.T. et al. A multi-material platform for imaging of single cell-cell junctions under tensile load fabricated with two-photon polymerization. Biomed Microdevices 24, 33 (2022). https://doi.org/10.1007/s10544-022-00633-z

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