What is Flow Cytometry?
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 one or more laser beams. The particle’s relative size, relative internal complexity or granularity and relative fluorescence intensity are the properties measured by a flow cytometer. These characteristics are determined using an optical-to-electronic coupling system that records how the particle scatters incident laser light and emits fluorescence. Once the data is collected, it can be displayed as graphics or histograms and statistics can be generated.
Common applications of flow cytometry are assessment of cell surface receptor expression (immunophenotyping), proliferation studies (CFSE), intracellular cytokine analysis, detection of fluorescent proteins like GFP, phospho protein analysis, cell cycle and apoptosis studies.
In most applications, after the data is collected, the cells are sent to waste. Cell sorting allows us to capture and collect the cells of interest for further analysis. Once collected, the cells can be analyzed microscopically, biochemically or functionally.
What is the purpose of the Immunophenotyping Core?
The purpose of the platform is to offer, for a fee, a variety of flow cytometry services to the research institute community as well as researchers from other affiliation. Also, since the flow cytometry field is constantly changing, the platform has for goal to give its users access to the newest technologies and applications in the field.
What services does the core offer?
The core offers cell analysis and cell sorting services. All services are offered on an hourly rate. The fee varies depending on the service (cell sorting is more expensive then cell analysis) and the researchers affiliation. Free access to a computer equipped with the analysis program FlowJo is also offered to the platform’s users.
Cell Analysis Service:
The core is equipped with 2 cytometers used for cell analysis: a BD LSRFortessa and a BD FACSCanto II. To be allowed to use one of those 2 instruments, users must follow a 3 hours training course with the person in charge of the platform. Once trained, the users are allowed to acquire their samples by themselves. The platform also offers the option of asking the operator to do the analysis but it is more expensive. The users also have access to an analysis computer. This computer is equipped with FlowJo, a well knowed data analysis program. A one hour training course on this program is also available.
Cell Sorting Service:
The platform offers cell sorting services. The BD FACSAria is a high-speed sorter with a fixed-alignment cuvette flow cell. This flexible cytometer provides superior performance in high-speed cell sorting and multicolour analysis. Our Aria is able to detect up to 7 colors on 2 lasers (blue and red). The sorter is also equipped with an ACDU device, allowing single cell sorting in plates, ideal for cloning. Sorted cells can be put back in culture after the procedure, use in adoptive transfers or used for RNA extraction for example.