Hands-on Flow Cytometry Training
The Cytometry core is now offering a new Flow Cytometry Short course for all new users once every 2-3 weeks on a rolling schedule. This course is required for all new flow core lab users, and recommended for everyone else. It will cover the following foundational concepts and important information and offer real hands on practice with a real experiment of a 6 color panel of stained PBMCs.
The focus of this course is to teach the best practices when using this cutting edge technology to produce the most reproducible publication quality data for your research studies. We will cover the basics of instrument operations to proper controls, experimental set up and design as well as hands-on techniques in proper instrument operations, as well as the acquisition of a stained 6 color PBMC panel. Following the acquisition of real samples we will practice proper compensation set up as well as manual vs. digital compensation in your experiments. All of these topics and skills are required for new as well as experienced users to learn correctly and to apply them correctly to your daily research. Course Contents
- Introduction to Flow Cytometry instrumentation, Fluidics, Optics and Electronics
- Proper voltage set up on the instruments and why this is important
- Understanding Staining index and Titration MFI curves
- Proper use of Isotype, Unstained, Positive and Negative controls
- Proper use of FC antibody block to prevent non-specific staining
- Proper Compensation and FMO gating controls
- Panel design titrations, validation and optimization best practices
- Flow Cytometer Setup (BD LSRII BD CANTO II or BD LSRFortessa with FACSDiva v8) with special emphasis on digital vs manual compensation.
- Basics on flow cytometer settings – voltages, gating, dump channels, exclusion, and compensation.
- Hands-on acquisition of Human Peripheral Blood Mononuclear Cells (PBMC) with a basic flow panel (6 colors). 2 teams to acquire 2 different panels.
- Each attendee will setup a flow cytometer and perform manual compensation, acquire data and discuss data analysis as a team in Flowjo or FCS express 6.0 software
The course costs 125.00 dollars. For more information please contact ICBR Cytometry core at ICBR-Cytometry@ad.ufl.edu