A multiplexed tissue imaging platform @Newcastle University for mapping cell types, states and interactions in human development, health and disease

Cells are the building blocks of life. The human body is thought to contain around 37.2 trillion cells and each one plays a role in determining our health and longevity. Different cell types have specialised functions such as muscle cells that allow us to move, brain cells that allow us to control our bodies, and immune cells that fight infection. In certain situations, even one cell behaving in an abnormal "anti-social" way in the body can lead to catastrophic outcomes such as cancer, neurodegeneration and auto-immunity; where the body attacks itself. Over the last few years, technologies that can measure multiple properties of single cells have been instrumental in advancing our understanding of how individual cells and communities/structures of cells behave both in normal and abnormal conditions.
The word "cytometry" literally translates to "Cell Measurement" from the Greek words "Kytos" meaning "cell" and "metria" meaning "to measure". When my daughter was 8 years old, she asked me what I do for a job. I told her I was a "cytometrist" and it was my job to "interrogate" cells to find out what they have been doing and whether that had done something bad. She thought about this and replied, "That means you are a Cell Detective Daddy". Since then this analogy has really stuck with me as it explains what I do perfectly. I use various technologies to interrogate large populations of cells and try to find my list of suspects in a disease (crime).
Until recently, it was only possible to ask many questions of our cells using so called "suspension" technologies where the cells had to be in a liquid. This was fine for things like blood samples, but not for situations where we wanted to measure cells in tissue, as it required us to break the structures down and release the individual cells for analysis. There are several major issues with this 1) It can kill certain or all cells in the sample, 2) It can change the cells in the sample making them gain or lose expression of important proteins/genes thus disguising their identity, 3) You lose the spatial location of the cell in the tissue with respect to its neighbours making it near impossible to now place the suspect cell at the crime scene.
The power of the technology we are requesting will allow us to do just that. It will have the capability to ask as many if not more questions of cells in the tissue space as we could in suspension, but importantly preserve the spatial context and cellular neighbourhoods. To use another analogy, suspension technologies are like the board game "Guess Who", but spatial technologies are more like the game "Cluedo". We can find both the suspect and place then at the crime scene; this is utterly essential if we want to solve serious and pressing issue sin human health and disease. The platform we are requesting is cutting-edge and will be one of only two in the UK. It will be supported technical specialists within a very experienced, core facility and will help to develop their careers. This is very much in line with the UK government Science Council's "technician commitment" and an urgent need to retain, develop and increase the number of technicians within academic and industrial settings.
We will use this technology to interrogate cells across numerous human diseases as well as different developmental stages from Oocyte to foetus. The data will have a profound impact on our understanding of human health, development and disease and help to uncover possible targets for treatment.

This proposal seeks funds to purchase a highly multiplexed tissue imaging system that can measure over 250 protein biomarkers on fixed-formalin paraffin embedded (FFPE) or fixed-frozen tissue sections using a cyclical, fluorescent antibody approach. Until relatively recently, highly multiplexed measurements on cells within tissue samples was largely impossible due to various technical constraints requiring disaggregation to single cell suspension followed by approaches such as flow cytometry, suspension mass cytometry and single cell RNA sequencing. While these approaches allow for thousands of parameters to be measured across millions of cells, all spatial contact and neighbourhood information is lost and cellular composition and identity can change. Recent advances in imaging technology that include moving away from fluorescence detection to using different probes such as metal tags in combination with Imaging Mass Cytometry (IMC) expanded the number of antibody-based measurements on tissue sections to a maximum of 40 targets. However, IMC lacks the sensitivity of fluorescence detection and will never exceed 40 parameters due to the limits of available stable metal isotopes for antibody tagging. It also lacks the ability to perform mRNA or multi-omics-based measurements. The recent development of imaging systems that can perform near multiple cyclical fluorescence-based measurements on tissue using combinations of antibody (protein) or oligo (mRNA) probes has expanded the possible parameter space to near limitless numbers. This innovative technology will enable numerous researchers to address fundamental questions in both human health, disease and development. All the areas of research mentioned in the case for support will benefit from using such a technology and are strongly aligned to the UKRI-MRC delivery plan including; precision medicine, discovery science, advanced therapies, molecular and cellular medicine and multi-morbidities.