The Flow Cytometry Service at LSTM is open to all users from academia or SMEs (small and medium-sized enterprises) and Industry. Our service also provides fluorescence activated cell sorting (FACS) by highly trained operators.
The Flow Cytometry Service offers assistance in a variety of areas, including access to a Cytek Aurora and FACSARIA, training, experimental design, panel development and optimization, technical support and data analysis.
- Acquisition and sorting of cells (including HG3 organisms)
- Data acquisition
- Data analysis
- Training and advice on sample preparation and acquisition
- User assistance and machine troubleshooting;
- Help with design of experiments, multiparametric panels and sourcing of reagents
We have one cytometer and one cell sorter equipped with 488 (blue), 633 (red) and 405nm (violet) lasers allowing multiparametric analysis for more than 24 colours (Cytek Aurora cytometer) and up to 18 colours (BD Facs Aria III Cell Sorter).
BD FacsAria III Cell Sorter. This equipment is in a HG3 laboratory and can be used to sort several cell types and organisms including HG3 pathogens such as TB. Four way sorting is possible, tube holders include sizes from microtubes to 15mL tubes and collection is also possible on a 384-well plate (most of users use 96 well plate) and slides. Sample and sort collection tubes can be maintained at a cooling or heating temperature. The acquisition workstation for this instrument is a PC with BD FacsDiva 8.1 software. The FCS 3.0 data files can be exported and analyzed using other flow cytometry software applications. Post-acquisition compensation are allowed and this cytometer offer bi-exponential scales.
The Cytek Aurora is a flexible, easy to use spectral flow cytometer, offering the ability to run high complexity assays using a simplified workflow giving high fidelity data. The unique optics of the Aurora offer a one solution approach using its standardised optical bench to sample light from 365nm to 830nm for multiple laser excitation lines to spectrally sample fluorophores allowing for better discrimination of these dyes. This “spectral approach” requires no optical configuration adjustments by the operator and allows for the use of a broader range of dyes when compared to conventual flow. The Aurora offers unprecedented fluorochrome flexibility allowing for the use of highly overlapping dyes such as APC and AF6
The Aurora offers:
- The ability to remove autofluorescence in highly auto fluorescent tissue to and to use this a s a parameter to improve antigen resolution.
- The ability to consolidate panels through using highly overlapping dyes to accurately discern subsets in high density panels more than 24 colours.
- The measurement of SSC off the violet and blue laser lines provide high sensitivity to resolve particles such as EVs nearing 100nm.”
- 47 in conjunction enabling panels more than 24 colours to be achieve don a 3 laser (Violet, Blue and Red) configuration. This provides researchers with greater choice in reagent options and allowing for larger panels to be formed, reducing sample burden, research costs per assay and increasing research power.
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