Deconstructing body axis morphogenesis in zebrafish embryos using robot-assisted tissue micromanipulation.

Classic microsurgical techniques, such as those used in the early 1900s by Mangold and Spemann, have been instrumental in advancing our understanding of embryonic development. However, these techniques are highly specialized, leading to issues of inter-operator variability. Here we introduce a user-friendly robotic microsurgery platform that allows precise mechanical manipulation of soft tissues in zebrafish embryos. Using our platform, we reproducibly targeted precise regions of tail explants, and quantified the response in real-time by following notochord and presomitic mesoderm (PSM) morphogenesis and segmentation clock dynamics during vertebrate anteroposterior axis elongation. We find an extension force generated through the posterior notochord that is strong enough to buckle the structure. Our data suggest that this force generates a unidirectional notochord extension towards the tailbud because PSM tissue around the posterior notochord does not let it slide anteriorly. These results complement existing biomechanical models of axis elongation, revealing a critical coupling between the posterior notochord, the tailbud, and the PSM, and show that somite patterning is robust against structural perturbations.

Correction: Actuated 3D microgels for single cell mechanobiology.

Correction for 'Actuated 3D microgels for single cell mechanobiology' by Berna Özkale et al., Lab Chip, 2022, 22, 1962-1970, https://doi.org/10.1039/D2LC00203E.

Actuated 3D microgels for single cell mechanobiology.

We present a new cell culture technology for large-scale mechanobiology studies capable of generating and applying optically controlled uniform compression on single cells in 3D. Mesenchymal stem cells (MSCs) are individually encapsulated inside an optically triggered nanoactuator-alginate hybrid biomaterial using microfluidics, and the encapsulating network isotropically compresses the cell upon activation by light. The favorable biomolecular properties of alginate allow cell culture in vitro up to a week. The mechanically active microgels are capable of generating up to 15% compressive strain and forces reaching 400 nN. As a proof of concept, we demonstrate the use of the mechanically active cell culture system in mechanobiology by subjecting singly encapsulated MSCs to optically generated isotropic compression and monitoring changes in intracellular calcium intensity.

Tissue Engineering with Mechanically Induced Solid-Fluid Transitions.

Epithelia are contiguous sheets of cells that stabilize the shape of internal organs and support their structure by covering their surfaces. They acquire diverse morphological forms appropriate for their specific functions during embryonic development, such as the kidney tubules and the complex branching structures found in the lung. The maintenance of epithelial morphogenesis and homeostasis is controlled by their remarkable mechanics-epithelia can become elastic, plastic, and viscous by actively remodeling cell-cell junctions and modulating the distribution of local stresses. Microfabrication, finite element modelling, light-sheet microscopy, and robotic micromanipulation are used to show that collagen gels covered with an epithelial skin serve as shape-programmable soft matter. The process involves solid to fluid transitions induced by mechanical perturbations, generates spatially distributed surface stresses at tissue interfaces, and is amenable to both additive and subtractive manufacturing techniques. The robustness and versatility of this strategy for engineering designer tissues is demonstrated by directing the morphogenesis of a variety of molded, carved, and assembled forms from the base material. The results provide insight into the active mechanical properties of the epithelia and establish methods for engineering tissues with sustainable architectures.

Investigating Tissue Mechanics in vitro Using Untethered Soft Robotic Microdevices.

This paper presents the design, fabrication, and operation of a soft robotic compression device that is remotely powered by laser illumination. We combined the rapid and wireless response of hybrid nanomaterials with state-of-the-art microengineering techniques to develop machinery that can apply physiologically relevant mechanical loading. The passive hydrogel structures that constitute the compliant skeleton of the machines were fabricated using single-step in situ polymerization process and directly incorporated around the actuators without further assembly steps. Experimentally validated computational models guided the design of the compression mechanism. We incorporated a cantilever beam to the prototype for life-time monitoring of mechanical properties of cell clusters on optical microscopes. The mechanical and biochemical compatibility of the chosen materials with living cells together with the on-site manufacturing process enable seamless interfacing of soft robotic devices with biological specimen.

Modular soft robotic microdevices for dexterous biomanipulation.

We present a methodology for building biologically inspired, soft microelectromechanical systems (MEMS) devices. Our strategy combines several advanced techniques including programmable colloidal self-assembly, light-harvesting with plasmonic nanotransducers, and in situ polymerization of compliant hydrogel mechanisms. We synthesize optomechanical microactuators using a template-assisted microfluidic approach in which gold nanorods coated with thermoresponsive poly(N-isopropylmethacrylamide) (pNIPMAM) polymer function as nanoscale building blocks. The resulting microactuators exhibit mechanical properties (4.8 ± 2.1 kPa stiffness) and performance metrics (relative stroke up to 0.3 and stress up to 10 kPa) that are comparable to that of bioengineered muscular constructs. Near-infrared (NIR) laser illumination provides effective spatiotemporal control over actuation (sub-micron spatial resolution at millisecond temporal resolution). Spatially modulated hydrogel photolithography guided by an experimentally validated finite element-based design methodology allows construction of compliant poly(ethylene glycol) diacrylate (PEGDA) mechanisms around the microactuators. We demonstrate the versatility of our approach by manufacturing a diverse array of microdevices including lever arms, continuum microrobots, and dexterous microgrippers. We present a microscale compression device that is developed for mechanical testing of three-dimensional biological samples such as spheroids under physiological conditions.