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.

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.

Publications

10 25 50
 
Description Member of the BBSRC People and Talent Strategy Advisory Panel RTP Subgroup
Geographic Reach National 
Policy Influence Type Participation in a guidance/advisory committee
 
Description MITalent Programme
Amount £739 (GBP)
Organisation United Kingdom Research and Innovation 
Department Research England
Sector Public
Country United Kingdom
Start 08/2023 
End 09/2023
 
Title New Imaging Flow Cytometry technology 
Description We have carried out a competitive tender for the purchase of an imaging flow cytometer. This was awarded on the 2nd October 2023 to Cytex for an ImageStreamX MkII system with INSPIRE and IDEAS software, Machine Learning module, 2 additional years service contract. The instrument was installed on the 21st/22nd December 2023 and training took place on the 24th /25th January 2024. This is a completely new capability for the University of Warwick. 
Type Of Material Improvements to research infrastructure 
Year Produced 2024 
Provided To Others? Yes  
Impact Since the training our flow technical specialist, Dr Steven Servin, has been gaining experience on the instrument and the data analysis. As part of exposing users of our facility to the new instrument, he has been running their samples in parallel on the ImageStream so that they get an idea of what is possible with the new instrument. This is completely new capability and we are engaging new users. We are aware of future grant submissions that will use the instrument. 
URL https://warwick.ac.uk/fac/sci/lifesci/tic/flow
 
Title Research technical professional training 
Description In September 2023, our flow technical specialist Dr Steven Servin, took part in a shadowing opportunity at Newcastle Fow Cytometry Core Facility under Prof Andrew Filby, funded by MITalent. 
Type Of Material Improvements to research infrastructure 
Year Produced 2024 
Provided To Others? Yes  
Impact Steven attended a 2 day ImageStream User meeting and 2 days shadowing the ImageStream instrument at the University of Newcastle. Steven brought the knowledge and skills back to the University of Warwick, enabling him to engage new users in the use of the instrument at Warwick. 
 
Description Tour to industry 
Form Of Engagement Activity Participation in an open day or visit at my research institution
Part Of Official Scheme? No
Geographic Reach Regional
Primary Audience Industry/Business
Results and Impact One of the Industrial users of our facility, NanoSyrinx, requested that we provide a tour of our flow facility to the BIA Manufacturing Advisory Committee Leadership Programme (BIA MAC LeaP). This included representatives from different manufacturing biotechs (10 attendees) and provided exposure to the technologies at the University of Warwick and potential knowledge exchange.
Year(s) Of Engagement Activity 2024