21 March 2019
State-of-the-art technology has been vital to our ground-breaking research and discoveries over the years. Ensuring this technology is supported by world-class expertise is equally important.
It's the rationale behind a three day cytometry training workshop run at the Malaghan Institute this week by Dr Maria Jaimes and Janelle Shook from Cytek Biosciences, USA.
“With any tool, the more adept you are at using it the greater the value you gain from it,” says the Malaghan Institute's Head of Research Technology Kylie Price.
Biomedical research is no different. Often, teasing out clues and vital understanding on how the immune system works comes down to identifying a single cell or molecule out of millions of others – a difficult task. Therefore, it's essential that our scientists and technologists have the skills necessary to get the most value out of the cutting-edge technology housed at the Malaghan Institute. With that in mind, the primary goal for this intensive workshop was to help Malaghan scientists improve and refine their experiments when using cytometry.
Cytometry is an advanced type of analytical technique used at the Malaghan Institute to give vital information about cells for health research and diagnostics. In 2017, the Institute became one of the first laboratories in the world to adopt spectral cytometry with the acquisition of a Cytek Aurora, thanks to support from the Hugh Green Foundation.
“The Malaghan Institute scientists are quickly becoming leaders in the application of this technology,” says Kylie.
“Spectral cytometry enables us to deeply interrogate which cell populations are present and what these cells are doing in the context of the diseases we study. With advances in this technology we’ll be getting unprecedented amount of information from each precious sample.
“This workshop comes at the perfect time as we have now had the Auroras for over a year and while we are very comfortable designing and using the technology, Dr Maria Jaimes and Janelle Shook from Cytek have really honed our expertise and taken us to the next level – allowing us to live up to our name as a Cytek Centre of Excellence.”
During the workshop we had the opportunity to sit down with Dr Jaime, to ask her about the ins and outs of this ground-breaking technology:
What is cytometry and how does it work?
Flow = in suspension, liquid
Cyto = cells
Flow cytometry is a technology that simultaneously measures and then analyses multiple physical characteristics of single particles, usually cells, as they flow in a fluid stream through a beam of light. The properties measured include a particle’s relative size, relative granularity or internal complexity, and relative fluorescence intensity. These characteristics are determined using an optical-to-electronic coupling system that records how the cell or particle scatters incident laser light and emits fluorescence
What is the difference between spectral cytometry and conventional cytometry?
"Spectral cytometry differs from conventional cytometry in its overall optical design and how the data is processed. In spectral systems, light is collected across the full spectrum of light, hence all detectors are used to collect measurements for every dye used on the system. In conventional systems, light for each dye is collected in a single detector in a narrow band of light, not over the full spectrum. These optical design differences require different methods of processing the data. In spectral cytometry, samples stained with multiple dyes are unmixed, or deconvolved, using a multiple least squares fitting approach to determine which dyes are present on each cell that passes through the system by comparing the unknown cell signals to an established set of controls for each dye that could be on the cell. In conventional cytometry, compensation is used to subtract signal that spills over into the detectors where that dye does not belong to correct the data so only signal from the dye matched to that detector appears in the final data."
"Spectral cytometry is a ground-breaking tool for researchers because it provides them more flexibility in building and expanding panels, eliminates the need to change hardware configurations so a variety of panels are easily accommodated, and allows researchers to extract autofluorescence, hence making highly autofluorescent cell types easier to study with spectral flow."
What's the future for this technology?
"We see this technology extending the boundary of what is possible to achieve in mulitcolour panels and improving the quality of published flow cytometry data with the system’s high sensitivity," says Dr Jaimes. "We plan to expand the capabilities of the current technology by adding additional lasers and expanding detector arrays to accommodate new near IR dyes, and in the future incorporate this technology into new cell analysis products."