Multi-modal high throughput live cell system to track biomechanical properties, coupled with fluorescence imaging

Our body is made up of tissues and organs that are mostly expandable and stretchy. Even structural tissues like tendons and bones have a degree of flexibility, and it is crucial for these tissues to be able to return to its original state after pressure, otherwise they will be injured and lose their function. There are different types of stretchiness, with two major types - elastic materials (like springs or rubber bands) can instantaneously recover its original shape after deformation, whereas viscoelastic materials (like jello) require a bit more time to regain its shape. Living tissues, such as skin, exhibit viscoelastic properties, which is why pressure marks (e.g. left by wristwatches) tend to take a while to disappear. Healthy tissues are also not uniform in its stretchiness, and abnormal changes in this stretchiness usually drives disease states like cancer and fibrosis. Thus it is crucial to understand how cells respond to changes in the stretchiness, in order to identify new therapeutics to stop abnormal cellular behaviour. It is also important for us to create a map of the elasticity/viscoelasticity across a tissue, so that we can create better materials to be used for replacement surgeries (e.g. hip replacement). Currently it is difficult to make these measurements using pre-existing technologies, as the existing instruments require very specific ways of preparing the samples, and take a long time for a measurement to be made. The purpose of this application is to buy a new machine known as the Pavone, which allows use to measure the different types of stretchiness quickly and efficiently. This also allows for biomaterials being developed at The University of Manchester to be tested for its suitability in supporting live cells, as the machine also supports long-term culture of cells and tissues. Researchers across the University will use the machine to understand fundamentally how cells respond to changes in the stretchiness of its environment, and screen for new biomaterials for tissue engineering. The machine can also be used to investigate new drugs that could be used to stop the abnormal changes in the stretchiness of the tissues, thus stopping disease progression.

The purpose of this proposal is to purchase a Pavone nanoindentation system with fluorescence imaging and environmental control modules. As the need for deeper understanding of how elasticity/viscoelasticity affects cellular behaviour both at the micro- and macro-scales arises, combined with the knowledge that complex biological systems have dynamic and reciprocal changes in biomechanics/cellular behaviour over time, this is an opportune moment to invest in a system that allows for real-time tandem measurements of both biomechanics and cellular behaviour through imaging. As of yet, none of the traditional microscopy/atomic force microscopy/nanoindentation instruments possess the capacity to do combinatorial long term fluorescence/brightfield live imaging and biomechanics measurements of multi-well plates. As such, users interested in correlating biomechanics with cellular phenotype and/or live tracking of changes in biomechanics have not been able to do so in a sophisticated manner. In addition, users interested in using current AFM/nanoindentation systems in quality control of their biomaterials have not been able to do so in a high throughput manner due to the specific nature of the vessels required for use on the systems. These bottlenecks can be addressed by the Pavone system, which will in turn free up capacity on the existing systems and allow them to be used for other experiments requiring more advanced/precise imaging or biomechanical measurements. This system is of huge interest to researcher not only in biological and medical sciences, but also to biofabrication and materials engineering, to ensure the materials made are biologically relevant and could sustain viable cells. This equipment will also benefit the wider research community beyond Manchester, driving future collaborations and research findings.