Imaging Flow Cytometry Instrument
Lead Research Organisation:
University of Warwick
Department Name: School of Life Sciences
Abstract
Flow cytometry and microscopy play an integral role in molecular science research. Flow cytometry is used to detect and measure fluorescently labelled characteristics of thousands of cells or particles over a timescale of seconds; microscopy is used to visualise cells and particles invisible to the eye on an individual basis. An imaging flow cytometer combines these two technologies, enabling fast visualisation of thousands of cells that can be used to quantify sub-cellular characteristics, removing the limitations of the separate techniques. This combined technology opens up an extensive range of novel applications.
At the University of Warwick, we have world-class research making a positive contribution to the future of our economy and our health. We will use the instrument for a broad range of research, from the fundamentals of how cells work and the mechanisms underpinning gene expression, to understanding reproduction in plants and improved crop yields. At a mechanistic level, we will improve our understanding in many areas, including the immune system, cell population heterogeneity, the genes involved in pregnancy, and processes underpinning antibiotic resistance.
We request funds for an Amnis ImageStreamX Mk II because it has the versatility to support cutting edge science in a broad range of research areas. For example, the range of magnifications available will ensure that we can analyse the variety of sample sizes anticipated (nanoparticles, yeast, bacteria, mammalian and plant cells). The instrument can also deliver up to 12 images (fluorescence, brightfield and darkfield) of every cell with speeds of up to 5000 cells per second. This will ensure that the instrument has the flexibility for an extensive range of applications.
The instrument will be established in the University of Warwick's Flow Cytometry Shared Resource laboratory, where it will be managed and supported by two research technical professionals. This environment will ensure that the instrument is well maintained and supported. The technical team will provide training, management and financial oversight and will ensure that use of the instrument incorporates sustainability practices, such as energy and consumable use. The instrument will be accessible to a wide userbase, both internal and external to the University, enhancing the bioscience research capabilities nationally.
At the University of Warwick, we have world-class research making a positive contribution to the future of our economy and our health. We will use the instrument for a broad range of research, from the fundamentals of how cells work and the mechanisms underpinning gene expression, to understanding reproduction in plants and improved crop yields. At a mechanistic level, we will improve our understanding in many areas, including the immune system, cell population heterogeneity, the genes involved in pregnancy, and processes underpinning antibiotic resistance.
We request funds for an Amnis ImageStreamX Mk II because it has the versatility to support cutting edge science in a broad range of research areas. For example, the range of magnifications available will ensure that we can analyse the variety of sample sizes anticipated (nanoparticles, yeast, bacteria, mammalian and plant cells). The instrument can also deliver up to 12 images (fluorescence, brightfield and darkfield) of every cell with speeds of up to 5000 cells per second. This will ensure that the instrument has the flexibility for an extensive range of applications.
The instrument will be established in the University of Warwick's Flow Cytometry Shared Resource laboratory, where it will be managed and supported by two research technical professionals. This environment will ensure that the instrument is well maintained and supported. The technical team will provide training, management and financial oversight and will ensure that use of the instrument incorporates sustainability practices, such as energy and consumable use. The instrument will be accessible to a wide userbase, both internal and external to the University, enhancing the bioscience research capabilities nationally.
Technical Summary
Research at the University of Warwick (UoW) has significant relevance to the BBSRC's strategic delivery plan and priorities. Our research will significantly benefit from the capabilities of an imaging flow cytometer, enabling statistical population analysis of characteristics such as particle morphology, co-localisation and signalling, at speeds of up to 5000 cells per second.
We are requesting funds for an Amnis ImageStreamX MkII imaging flow cytometer. This instrument combines multiparametric fluorescence measurements of a population with high-throughput brightfield, darkfield and fluorescent imaging of each cell and powerful data analysis software, IDEAS, to facilitate complex image analysis and machine learning. The instrument will be capable of:
- Detecting a broad range of fluorophores (five laser system, 375nm, 405nm, 488nm, 561nm and 642nm), complementing existing technology at the UoW
- A range of magnifications (60x/40x/20x) to ensure support for a broad range of sample types
- Resolving small particles, such as nanoparticles/extracellular vesicles and microplastics, using a high power 400mW blue laser, 60x magnification and high gain mode
- Capturing up to 12 high resolution images of every cell passing through the instrument
- Capturing all structures within the cell into one crisp plane of focus with an Extended Depth of Field (EDF) option
The instrument will be integrated into a specialised Flow Shared Resource Laboratory, where it will be professionally managed and supported for multi-user access, both internally and nationally. The instrument will be managed by the PI, an experienced research technology facility manager, and supported by the Research Co-I, a flow cytometry specialist. The research team Co-Is will support the development of research applications on the imaging flow cytometer, sharing these with the research community and building on the UK's bioscience capabilities.
We are requesting funds for an Amnis ImageStreamX MkII imaging flow cytometer. This instrument combines multiparametric fluorescence measurements of a population with high-throughput brightfield, darkfield and fluorescent imaging of each cell and powerful data analysis software, IDEAS, to facilitate complex image analysis and machine learning. The instrument will be capable of:
- Detecting a broad range of fluorophores (five laser system, 375nm, 405nm, 488nm, 561nm and 642nm), complementing existing technology at the UoW
- A range of magnifications (60x/40x/20x) to ensure support for a broad range of sample types
- Resolving small particles, such as nanoparticles/extracellular vesicles and microplastics, using a high power 400mW blue laser, 60x magnification and high gain mode
- Capturing up to 12 high resolution images of every cell passing through the instrument
- Capturing all structures within the cell into one crisp plane of focus with an Extended Depth of Field (EDF) option
The instrument will be integrated into a specialised Flow Shared Resource Laboratory, where it will be professionally managed and supported for multi-user access, both internally and nationally. The instrument will be managed by the PI, an experienced research technology facility manager, and supported by the Research Co-I, a flow cytometry specialist. The research team Co-Is will support the development of research applications on the imaging flow cytometer, sharing these with the research community and building on the UK's bioscience capabilities.
