Using super-resolution live cell imaging to understand the dynamics of disease
Lead Research Organisation:
University of Cambridge
Department Name: Cambridge Institute for Medical Research
Abstract
Humans are made up of trillions of individual cells which grow, divide and communicate with each other in tightly controlled ways. Diseases are often caused by errors in fundamental processes that happen inside our cells and understanding both these processes themselves and how they go wrong in inherited or infectious diseases can be critical to developing new drugs and diagnostics.
Understanding how cells function depends heavily on microscopes, which can visualise compartments (known as organelles) within cells at high resolution, even though they are less than 0.001mm in size. Some microscopes can do so while the cells are living and growing, allowing scientists to monitor cellular processes in real time as they are happening. However, microscopes are often highly specialised, useful for certain experiments but not others. We are requesting funding for a new microscope (a Zeiss LSM980/Airyscan2) that will be based in an already existing core facility in the Cambridge Institute for Medical Research (CIMR), and used by researchers in Cambridge, specifically CIMR, the MRC Mitochondrial Biology Unit and the UK Dementia Research Institute Cambridge Centre. Across the three institutes there are 40 research groups and >300 scientists who work on understanding how defects in cells cause human disease. Almost all of these researchers use microscopy as part of their work, and the core facility is therefore very busy. Purchasing a new microscope is important because two of our most highly used microscopes are old, and parts for them will soon no longer be available, meaning they won't be able to be repaired. However, the new microscope is much more than a replacement as it includes new hardware features that radically improve resolution and sensitivity and allow users to switch between different modes and different kinds of microscopy, a feature that is unique to this machine. It also has new software, including artificial intelligence, which allow the generation of much larger quantities of data because certain steps of operating the microscope can be automated.
Scientists will use the microscope on a range of different research projects. In some projects they will study organisms that infect human cells (such as those that cause malaria, or a highly understudied disease called scrub typhus), in order to understand how they manipulate cellular processes, and whether these host-pathogen interactions can be targeted by drugs. In others they will use live microscopy to study how proteins move between organelles within human cells, or how organelles come into contact with each other, and see how mutations disrupt these processes. Errors in these pathways are often associated with severe disease, such as neurological or mitochondrial disease, and understanding these molecular details can help the development of new therapeutic approaches. In all cases, the unique technical features of the LSM980/Airyscan2 will allow researchers to carry out experiments that would simply not be possible without it.
The new microscope will be used primarily by researchers in the three institutes, which are situated in two buildings immediately next to each other within the University of Cambridge School of Clinical Medicine, but it will also be available to any researcher who wants to use it, including those from institutes and universities outside Cambridge. All users will pay for using the machine, which will help to maintain it and keep it running optimally for many years. It will be managed and run by a highly skilled and experienced team who have been running the CIMR Microscopy Facility for >20 years. These outstanding scientists work very closely with the research teams to help them design and deliver their experiments, a collaborative and supportive approach that makes the science better.
Understanding how cells function depends heavily on microscopes, which can visualise compartments (known as organelles) within cells at high resolution, even though they are less than 0.001mm in size. Some microscopes can do so while the cells are living and growing, allowing scientists to monitor cellular processes in real time as they are happening. However, microscopes are often highly specialised, useful for certain experiments but not others. We are requesting funding for a new microscope (a Zeiss LSM980/Airyscan2) that will be based in an already existing core facility in the Cambridge Institute for Medical Research (CIMR), and used by researchers in Cambridge, specifically CIMR, the MRC Mitochondrial Biology Unit and the UK Dementia Research Institute Cambridge Centre. Across the three institutes there are 40 research groups and >300 scientists who work on understanding how defects in cells cause human disease. Almost all of these researchers use microscopy as part of their work, and the core facility is therefore very busy. Purchasing a new microscope is important because two of our most highly used microscopes are old, and parts for them will soon no longer be available, meaning they won't be able to be repaired. However, the new microscope is much more than a replacement as it includes new hardware features that radically improve resolution and sensitivity and allow users to switch between different modes and different kinds of microscopy, a feature that is unique to this machine. It also has new software, including artificial intelligence, which allow the generation of much larger quantities of data because certain steps of operating the microscope can be automated.
Scientists will use the microscope on a range of different research projects. In some projects they will study organisms that infect human cells (such as those that cause malaria, or a highly understudied disease called scrub typhus), in order to understand how they manipulate cellular processes, and whether these host-pathogen interactions can be targeted by drugs. In others they will use live microscopy to study how proteins move between organelles within human cells, or how organelles come into contact with each other, and see how mutations disrupt these processes. Errors in these pathways are often associated with severe disease, such as neurological or mitochondrial disease, and understanding these molecular details can help the development of new therapeutic approaches. In all cases, the unique technical features of the LSM980/Airyscan2 will allow researchers to carry out experiments that would simply not be possible without it.
The new microscope will be used primarily by researchers in the three institutes, which are situated in two buildings immediately next to each other within the University of Cambridge School of Clinical Medicine, but it will also be available to any researcher who wants to use it, including those from institutes and universities outside Cambridge. All users will pay for using the machine, which will help to maintain it and keep it running optimally for many years. It will be managed and run by a highly skilled and experienced team who have been running the CIMR Microscopy Facility for >20 years. These outstanding scientists work very closely with the research teams to help them design and deliver their experiments, a collaborative and supportive approach that makes the science better.
Technical Summary
Fundamental cellular processes - membrane trafficking and homeostasis of proteins and organelles can be disrupted by genetic variation or manipulated by intracellular pathogens, leading to human disease. Understanding the detailed impact of these changes at a cellular level is essential to the development of new disease models, diagnostics and drugs, and is dependent on high-end microscopy. This application will support the purchase of a Zeiss LSM980/Airyscan2 microscope which fits into a unique niche combining key advantages of confocal, super-resolution and high content screening microscopy. The hardware features give it significant increases in resolution and sensitivity, as well as the flexibility to combine super-resolution and photomanipulation on the same instrument. Software features including AI, direct processing and real-time deconvolution provide significant advantages in increasing automation and throughput without sacrificing resolution.
The microscope will be used by researchers across the Cambridge Institute for Medical Research, MRC Mitochondrial Biology Unit and UK Dementia Research Institute at Cambridge for a range of MRC-funded and/or relevant projects in molecular and cellular medicine, infection and immunity, neurosciences and translational research. It will enable researchers to perform live cell microscopy at a new level of throughput and resolution to better understand dynamic changes in organelle organisation and protein movement within and between organelles and how that is affected by disease-causing mutations. The increased sensitivity will allow them to follow organelle dynamics within small intracellular pathogens and visualise proteins at endogenous levels, removing the need for overexpression that can sometimes lead to mislocalisation. The microscope will be situated in a well-established core facility where it will be supported by a highly experienced and collaborative technical team, underwritten by extensive in-kind support.
The microscope will be used by researchers across the Cambridge Institute for Medical Research, MRC Mitochondrial Biology Unit and UK Dementia Research Institute at Cambridge for a range of MRC-funded and/or relevant projects in molecular and cellular medicine, infection and immunity, neurosciences and translational research. It will enable researchers to perform live cell microscopy at a new level of throughput and resolution to better understand dynamic changes in organelle organisation and protein movement within and between organelles and how that is affected by disease-causing mutations. The increased sensitivity will allow them to follow organelle dynamics within small intracellular pathogens and visualise proteins at endogenous levels, removing the need for overexpression that can sometimes lead to mislocalisation. The microscope will be situated in a well-established core facility where it will be supported by a highly experienced and collaborative technical team, underwritten by extensive in-kind support.
