Flow cytometry: analysis and sorting cells
- Duration 2 day – 12 hours Doctoral School equivalent
- Language English or French depending on audience
- Name of the teachers Gaël Brasseur
- Administrative head Ben Field and Mila Sirinelli
- Number of students 2 to 5
This workshop will allow to analyse single cells from mixed or homogenous prokaryotic or eukaryotic samples for their size, diffraction multi-fluorescences and distribution of the population with proper statistics. Participants will learn theoretical and practical aspects of flow cytometry.
The Plinius Cursus offers the “Flow cytometry” workshop which will allow to understand the possibilities to analyse cells at single level in order to estimate the distribution of a population with its accurate statistics for the different parameters observed. This will also allow to sort physically the cells on different samples based on chosen characteristics (size, fluorescence etc.).
At the end of the workshop, attendees will be able to understand the multiple possibilities given by the analysis and sorting samples by flow cytometry. Attendees will know how to design an experiment, how to prepare and to run samples, the gating strategy, to handle the statistics and data during experiments or in a post-acquisition step with specific softwares. Attendees will get a better idea of the diversity of projects and measurements using a flow cytometer and also its limitations.
Timeline
- Day 1: Theory: Introduction to the theory of flow cytometry : the fluidic system, optical design of lasers and detection, single particle analysis, signal treatment, description of different type of instruments, for which samples, choice of chemical or protein-based fluorochromes, gating strategy, sorting samples, statistics of data.
- Day 2: Experiments : calibration of the instrument, Quality Control and drop-delay with beads, running mixed and single bacterial samples, gating strategy, analysis data, choosing a combination of parameters for sorting sample (size, diffraction, fluorescence or combined parameters) data processing in post-acquisition with Prosort and powerful softwares (ex. FlowJO).
Students prerequisites
- Agreement of the thesis director in case of PhD student
- Attendees can bring their own non pathogenic biological samples. For this purpose, it is necessary to contact Gaël Brasseur to determine the types of samples required.
Location
Campus J. Aiguier, Bat N, ground floor, Room N025A.
Labo impliqué dans la formation. LCB, team IAM.