Flow Cytometry Core Facility

Novel Method Development

Novel Method Development

Staff at Newcastle FCCF are engaged in areas of novel method development across a broad range of cytometry.

Label Free Cytometry

Science beyond the spectrum: using imaging cytometry and machine learning to phenotype cells based upon brightfield characteristics

Staff have been pursuing research into the identification of cellular subtypes without the use of antibodies or fluorophores. This area of work shows great promise, utilising the advancements in image capture and data analysis. Newcastle FCCF has been working in collaboration with researchers at Ghent University using images of white blood cells generated by the Amnis ImagestreamX and the Zeiss Celldiscoverer 7.

This work has already yeilded successful publications and has developed a successful international relationship between Ghent and Newcastle. 

Mass cytometry standardisation

Creating knowns in a field of unknown: new technology requires new standards, at Newcastle FCCF we are developing new standardisation for mass cytometry to add confidence and boost instrument optimisation

We are fortunate to have the Fluidigm Helios suspension mass cytometry instrument and the Hyperion imaging mass cytometry instrument allowing high dimensional analysis of cells in suspension and tissue sections on slides. As this new technology is rapidly utilised around the world, efforts by Newcastle FCCF have been made to create standardisation on both instruments.

As CyTOF uses metal conjugated antibodies and a time of flight mass spectrometer, much work is needed in standardisation of instrument sensitivity using all available rare earth metals.

When used in Hyperion mode, tissue sections are ablated using a high powered UV laser and the antibody-metal conjugates are detected using the same mass detection as the Helios. Here the challenge is different, with slower acquisition speeds and restriction of one slide per analysis, the staff at Newcastle FCCF have been working to improve control tissue staining for each ablation without the need to change slides. 


Sample barcoding

Small differences, big impacts: looking in depth at the effect of barcoding on kinetic flux assays in flow cytometry

Studying calcium flux using flow cytometry has been common place since the first published work on the subject in 1995. Protocols are now often published and tbhe assay is done frequently both at Newcastle FCCF and across the world.

Due to its kinetic nature, calcium flux assays can have a large amount of variation sample to sample. To counter this, work within Newcastle FCCF has been focused on tightening assay variation using sample barcoding and simultaneous acquisition and stimulation.

Genomic Cytometry

The power of combined technologies: Genomic cytometry brings the power of cell sorting and flow cytometry together with single cell sequencing.

The field of Genomic Cytometry is a rapidly expanding area of research. Gathering genomic and transcriptomic information on a single cell level is allowing scientists to accurately and quickly assess their cells of interest in huge detail, identifying never before seen subtypes and disease states. 

To assess rare cells it has become common practice to pre sort cells to enrich for a rare cell type prior to genomic analysis. Using the power of cell sorting, pure cells can be obtained and a more targeted approach to single cell genomics can be carried out. The FCCF has successfully worked with academics to produce high quality cells for single cell genomic analysis resulting in high impact publications. Proper handling of cells prior to sequencing is key to generating robust data. Come and dicuss your project needs with us and take advantage of our in depth knowledge of cell sorting prior to genomic analysis to get the best quality data possible. 

Staff at the FCCF have played a key role working with international labs and Becton Dickinson to develop the Rhapsody system

For your sequencing needs please contact the Genomics Core Facility at Newcastle University