Expanding multiparameter analyses of single cells and small particles with a 5-laser spectrally enabled flow cytometer
Lead Research Organisation:
University of Edinburgh
Department Name: Sch of Biological Sciences
Abstract
Investments in fundamental "discovery" science as it relates to our immune system have underpinned both the twentieth century "age of vaccines" and the ongoing twenty-first century revolution in the treatment of diseases such as cancer, arthritis and eczema, and are now paving the way for more effective vaccines and for new treatments for inflammatory bowel disease and type 1 diabetes. Advances in vaccine technology were essential in bringing the Covid pandemic to its end and novel "immunotherapies" are increasing life expectancy and quality of life for millions of people with chronic life-limiting diseases, including cancers which are inoperable or resistant to chemotherapy and radiotherapy.
Although investment in fundamental research, such as that described in this proposal, can take time to benefit society, understanding the cellular and molecular processes that comprise a healthy and functional immune system is essential if we are to continue to develop new drugs and treatments, control the spread of infectious diseases and remain healthy and productive throughout our ever-longer lives.
In order to study these processes in detail, researchers need to be able to extract as much information as possible from each biological sample they generate. The more detailed the information we can get, the better we will understand these processes. This, in turn, will allow us to develop more effective strategies (better therapeutic outcomes with fewer side effects, for a larger proportion of those affected) to enhance and sustain health during development, ageing and infection.
We are proposing to buy a new machine that will enable us to measure up to 50 different pieces of information per sample. By analysing multiple samples in parallel, we can obtain hundreds and thousands of pieces of information from a single person, animal or experiment. These samples are a precious resource. Maximising the amount of information we get from each one not only minimises the size and number of samples we need to take (which saves money and is more ethical) but, crucially, exponentially increases our levels of understanding.
Although investment in fundamental research, such as that described in this proposal, can take time to benefit society, understanding the cellular and molecular processes that comprise a healthy and functional immune system is essential if we are to continue to develop new drugs and treatments, control the spread of infectious diseases and remain healthy and productive throughout our ever-longer lives.
In order to study these processes in detail, researchers need to be able to extract as much information as possible from each biological sample they generate. The more detailed the information we can get, the better we will understand these processes. This, in turn, will allow us to develop more effective strategies (better therapeutic outcomes with fewer side effects, for a larger proportion of those affected) to enhance and sustain health during development, ageing and infection.
We are proposing to buy a new machine that will enable us to measure up to 50 different pieces of information per sample. By analysing multiple samples in parallel, we can obtain hundreds and thousands of pieces of information from a single person, animal or experiment. These samples are a precious resource. Maximising the amount of information we get from each one not only minimises the size and number of samples we need to take (which saves money and is more ethical) but, crucially, exponentially increases our levels of understanding.
Technical Summary
Flow cytometry is a technique to assess the phenotypic characteristics and functional properties of individual cells and particles by detecting spectral emissions of endogenously expressed (typically after genetic manipulation) or exogenously attached (via antibodies, dyes or cognate receptor-ligand interactions) fluorescent probes. The ability to capture and quantify multiple parameters on thousands of individual cells or particles in a single sample makes flow cytometry one of the most powerful platforms available for simultaneous quantitative and qualitative analysis of intact cells at scale. Recent technological advances and reagent development make it possible to simultaneously detect many more parameters than previously possible. These advances will facilitate the generation of unprecedented insights into immunology, parasitology and cell biology within the College of Science and Engineering, at UoE. The equipment we propose to purchase (a 5-laser spectral analyser) offers 3 advantages over existing flow cytometers: -
1) Simultaneous detection of up to 50 individual fluorescence parameters in a single sample. This would dramatically enhance capability within the SBS which currently stands at 16.
2) Identification and subtraction of autofluorescence, resulting in increased resolution of otherwise difficult to analyse samples such as macrophages and preparations from whole organisms (e.g. Drosophila spp).
3) Improved resolution of small particles such as extracellular vesicles, bacteria and yeast.
These improved capabilities will be exploited to increase our understanding of: -
i) composition, development and function of healthy cell populations
ii) immune responses during infection and disease
iii) heterogeneity of immune responses within and between populations (e.g. by age, sex, environment)
iv) the effects of environmental conditions on microorganisms, to explore cutting-edge biotechnologies (e.g. plastic-recycling and biomining).
1) Simultaneous detection of up to 50 individual fluorescence parameters in a single sample. This would dramatically enhance capability within the SBS which currently stands at 16.
2) Identification and subtraction of autofluorescence, resulting in increased resolution of otherwise difficult to analyse samples such as macrophages and preparations from whole organisms (e.g. Drosophila spp).
3) Improved resolution of small particles such as extracellular vesicles, bacteria and yeast.
These improved capabilities will be exploited to increase our understanding of: -
i) composition, development and function of healthy cell populations
ii) immune responses during infection and disease
iii) heterogeneity of immune responses within and between populations (e.g. by age, sex, environment)
iv) the effects of environmental conditions on microorganisms, to explore cutting-edge biotechnologies (e.g. plastic-recycling and biomining).
