Critical to investigating the complex immune response to disease is the capacity to measure cells at the single-cell level, examining multiple features of the cell simultaneously. However, the development and implimentation of new methods that can achieve this is inherently challenging. To address this, I have lead a number of developments of new cytometry technologies or assays.
High-dimensional cytometry and single-cell technologies
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Mass cytometry (CyTOF) In 2014, during my PhD in the laboratory of Prof. Nicholas King, I was the first to develop and apply assays using Australia's first mass cytometer (CyTOF), having recieved training in the laboratory of Dr. Garry Nolan at Stanford University. I used this novel technology to comprehensively profile inflammatory mobilisation of the murine haematopoietic system in the bone marrow during viral encephalitis (Ashhurst et al. 2019). This early work in mass cytometry lead to the publication of a book, featuring a collecting of up-to-date and cutting-edge protocols in mass cytometry, which is now a standard resource in the field (HM McGuire, TM Ashhurst (eds). 2019).
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High-dimensional flow cytometry In 2016, with the [Sydney Cytometry Core Research Facility](), I took a leading role in the development of a novel **high-dimensional flow cytometry** platform, our world first [10-laser 30-parameter ‘LSR-X’](https://sydneycytometry.org.au/hdflow) platform, developed in collaboration with BD. This was the first high-parameter cytometry to be implimented in Australia, and the 25-colour panel we developed for the system was the first published panel that reached above 18 colours ([Ashhurst, Smith, King. 2017](https://currentprotocols.onlinelibrary.wiley.com/doi/abs/10.1002/cpim.37)). These efforts lead to participation as a speaker in the ACS 'Polychromatic Flow Cytometry' roadshow alongside Dr. Pratip Chattopadhyay, teaching in multiple cities aross Australia and New Zealand in 2016.
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Spectral cytometry More recently, I have implemented high-dimensional **spectral cytometry** platforms and panels within the Sydney Cytometry Facility, and performed comprehensive evaluations of both conventional and spectral cytometry ([Niewold\*, Ashhurst\* et al. 2020](https://onlinelibrary.wiley.com/doi/abs/10.1002/cyto.a.24211)).
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Single-cell transcriptomics Most recently, I lead the establishment of the genomic/molecular cytometry arm of the Sydney Cytometry Facility, incorporating the BD Rhapsody, a **multiomic single-cell transcriptomic system**.
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High-dimensional imaging/spatial technologies
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Imaging Mass Cytometry (IMC) Recently, with the Sydney Cytometry Facility, I developed and applied the first **Imaging Mass Cytometry (IMC)** assays in Australia, to profile cellular infiltration into the brain of West Nile or Zika virus infected mice. This work has been presented at a number of recent international meetings (refs).
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