What is Flow Cytometry?
Flow cytometry is a generic name for techniques that involve the analysis and/or separation of cells, or other particles, by quantification of optical parameters. As widely practised today, this routinely uses suspensions of living, or fixed cells streamed past one or more lasers from which various light scattering and fluorescence parameters are detected using different detectors. Although these techniques are most widely used in mammalian biology, they can be applied to almost any particle with useful optical properties, including bacteria, viruses, subcellular organelles, and artificial particles, for example, polystyrene beads bearing ligands or antibodies.
Examples of applications
Cell sorting
- Separation of up to 6 sub-populations simultaneously.
- Bulk or single cell sorting into different collection tubes or multi-well plates (from 6-well to 1536-well plates)
- Imaging-enabled sorting using the CellView technology from the BD FACSDicover S8.
- Separation of live or fixed samples that can be used in several downstream applications like immunophenotyping, cell cycle analysis, western blots, CRISPR screening, SMART-Seq, 10x genomics, etc.
- The Cambridge Genomics Services facility is also located in the Department of Pathology and provides services like next-generation sequencing, microarray and analysis. Please contact them for more information.
Analysis
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Immunophenotyping
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Transfection efficiency by measuring fluorescent proteins levels
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Cell cycle
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Proliferation assays
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Apoptosis and cell viability assays
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RNA detection assays
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Intracellular calcium flux
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Rare event analysis
Instruments available in the facility
Cell Sorters
- BD FACSDiscover S8 with CellView technology (5 lasers, full spectrum flow cytometer, 78 algorithmically optimised fluorescent APDs covering the full spectrum. Imaging-enabled cell sorting using CellView)
- BD Aria III (4 lasers, conventional flow cytometer, 16 PMT detectors)
- BD Aria IIu (3 lasers, conventional flow cytometer, 9 PMT detectors)
The cell sorters can run samples safe to handle under containment levels 1 and 2 as per risk assessment. The Aria IIu is restricted to containment level 1.
Analysers
- Two Cytek Biosciences Aurora (5 lasers, full spectrum flow cytometer, 64 fluorescent APDs covering the full spectrum)
- Beckman Coulter CytoFLEX LX (5 lasers, conventional flow cytometer, 19 APD detectors)
- Beckman Coulter CytoFLEX S (4 lasers, conventional flow cytometer, 13 APD detectors)
- Thermo Fischer Attune NxT (4 lasers, conventional flow cytometer, 14 PMT detectors)
- BD LSR II (4 lasers, conventional flow cytometer, 17 PMT detectors)
The Attune NxT can run samples safe to handle under containment levels 1 and 2 as per risk assessment. All the other analysers are restricted to containment level 1 samples.
How to get access to the facility?
We welcome enquiries regarding possible uses.
All users who wish to use the flow cytometry facility located in the Department of Pathology need to request an account in our online booking system called PPMS.
Once the facility's manager approves your account, all sorting and training requests and bookings will be processed via PPMS.