Welcome to the MMRI Flow Cytometry Core
The core possesses a 4 laser, 16 channel BD FACS Symphony A3 flow cytometer with four laser lines 405nm, 488nm, 651nm and, 637nm and a 4 laser, 15 channel BD FACS Aria Fusion cell sorter with 405nm, 488nm, 561nm and 640nm laser lines. As well as software for the analysis of
The protocols on the Flow Core are basic protocols, they may need to be optimized for the users target populations and antibody panel.
The MMRI guide to flow cytometry is linked here and it provides background information on how flow cytometry works, what controls are appropriate and how to begin your experiments.
BD FACS Symphony A3
Our FACS Symphony A3 flow cytometer is suitable for a range of analysis from whole blood, immune organs, extracted leukocytes from organs such as the heart, phagocytosis assays, transfection efficiency and, confirmation of stem cell potential in iSPC cultures.
The core provides basic protocols which users can optimize for their own studies. These protocols include the staining of live cells, fixed cells, intracellular cytokine and transcription factor staining, cell cycle analysis and, cell tracking. Additionally, there is information on designing and building a flow cytometry panel and data acquisition and analysis that can be found here .
Users are required to contact Dr Samantha Le Sommer to arrange induction and training on the machine before you will be given access to the booking calendar. It is recommended that prior to buying in reagents/commencing work that you speak to the core to ensure your chosen panel is compatible with the FACS Symphony analyzer.
Details of the current laser, filter, and detector set up is below.
BS FACS Aria Fusion Cell Sorter
The BD FACS Aria Fusion sorter is capable of high-speed single cell sorting on both forward and side scatter plus up to 11 fluorochromes. Cells can either be sorted into 4-way tubes, or directly into plates or onto slides.
It is important to contact the core prior to any work to ensure that your cells are sortable on your current panel set up. Cells can only be sorted on surface markers, hence, for some populations sorting can be tricky. A set of considerations to take into account when planning your experiment can be found and basic protocols for the staining and sorting of cells can be found at this link here.
Please note that currently no solo users will be allowed unsupervised access to the cell sorter.
FlowJo Analysis Software
The core owns and maintains a license to the powerful analysis software FlowJo. This is installed on the core PC and is bookable via the Talen calendar. FlowJo is capable of both basic and advanced analysis on populations of interest, expression of markers, cell cycle analysis, proliferation and apoptosis. Training will depend on the user experiment, to arrange training on the software contact the core.