Developing a real-time live cell imaging platform at Newcastle University

Understanding how cells respond in real-time to their environment is a key observation in many disciplines of medical research. For example, understanding how cancer cells respond to novel drug treatments or how immune cells interact with other cell types in autoimmune disease. Simple incubator-based microscope-based platforms have existed for many years and have allowed us to monitor in real-time the behaviour of cells by following changes in: shape, division, migration and proliferation. Technology in this area has now vastly improved and combined with AI-driven image analysis algorithms, we are now able to study much more complex behaviours and with much greater throughput.

The Incucyte SX5 is a long-term live cell real-time microscopy platform that will enhance our research capabilities across a number of research fields at Newcastle University. By housing this platform within our established Bioimaging Unit, we can offer expert advice on experimental set up and data analysis and guarantee that the platform will be supported and used optimally throughout its operational lifespan. The ability of this machine to perform 6 independent studies concurrently provides superior multi-user capabilities over previous Incucyte generations which could only run one type of assay at a time. Research that will be carried out using this equipment includes the study of globally significant diseases where Newcastle University scientists are world-leaders these include: multiple cancer types (Newcastle Centre for Cancer), mitochondrial disorders (Wellcome Centre for Mitochondrial Research), respiratory, neurodegenerative and liver diseases. Using the Incucyte SX5, we not only carry out established and optimised experiments (using the previous-generation Incucyte Zoom technology) but will expand on them to incorporate complex kinetic measurements. For example, individual cellular analysis can now be performed which improves the accuracy of proliferative and migratory data. We can combine this with both drug and CRISPR screening to identify new therapeutic targets for disease. With the ability to detect up to 5 fluorescence colours the experimental capabilities are further enhanced. For example, cell cycle phase can be recorded in real-time, co-culture studies can be carried out such as visualisation and quantification of tumour-immune cell interactions to interrogate advanced therapeutic interventions. Excitingly, robust and consistent evaluation of 3D cell cultures is now possible and will be performed by a number of research groups which will increase the impact of their research programs and help deliver new disease models for their research field. With a strong neuroscience research base in Newcastle, there is a demand to access neuronal activity assays which only the SX5 model can offer. Similarly, ATP metabolism assays can be performed which are highly useful to our world-leading mitochondrial research teams.

In line with the principles developed by Newcastle University in its commitment to become carbon neutral by 2030, the cross-campus imaging platform developed will result in the removal, re-use (for spare parts) and recycling of existing older systems. It will also make use of existing incubators and require no further lab modifications or services. The platform will be networked to facilitate remote monitoring and experimental set-up. It will also reduce the requirement for staff to regularly transport across campus as well as duplicate experiments, media and reagents.

Understanding how cells respond to their environment in real-time is a vital experimental parameter in comparison to historic endpoint assays in a number of medical research disciplines. Long-term live cell imaging allows this to be addressed and with technology and software advances there are a number of kinetic assays which are now available. The objective of this proposal is to provide a platform within Newcastle University Bioimaging Unit which can deliver this type of data, support the users of the equipment with experimental design and analysis and enhance our research capabilities and capacity.

Over 30 research groups -with MRC strategic research priority interests- will, through the use of improved technology and a number of AI-driven analysis packages, exploit the real-time live-cell imaging capabilities. After the full evaluation of the Incucyte SX5 and other similar technologies and considering the level of demand, we determined that it is best in class and fully meets our needs. In particular it: is truly multi-user (6 concurrent experiment capacity), is fully customisable (independent, magnification, channel, observation frequency parameters can be set for each experiment) and has a suite of fully integrated AI-driven analysis modules. The system can also be accessed, and data shared remotely.
Research to be carried out using the SX5 will include: assessment of advanced disease models by organoid/spheroid analysis; high-throughput drug discovery experimentation in multiple disease models; fluorescence based assays to monitor cell cycle in real-time and co-culture studies to investigate cellular interaction. Enhanced capabilities will allow new assays to be developed including neurite outgrowth and metabolism (ATP assays).

Due to the plethora of possible assays, the number of research groups wishing to use this equipment, we anticipate this platform will support MRC-remit and impactful discovery science for years to come.