Flow Cytometry Basics: Self-paced e-learning courses from BD Biosciences. Focused on flow cytometry theory as employed in classical/conventional BD flow cytometers.
For Users of BD Instruments
FACSDiva YouTube Trainings: is the software that runs the and the . Each video is about 100 min long but goes through both the theory and the hands-on use of the software. These training videos were produced by Open Flow Cytometry, an open access Flow Cytometry education resource. The Open Flow initiative is a collaboration between the Francis Crick Institute (London, England), Memorial Sloan Kettering Cancer Center (NY, NY, USA) and the Whitehead Institute for Biomedical Research (Cambridge, MA, USA).
Part 1: Experimental Set-up in FACSDiva
Part 3: Compensation in FACSDiva
For Users of Instruments
Full Spectrum Flow Cytometry: After you have become familiar with basic flow cytometry theory, Open Flow Cytometry has released a training video on YouTube entitled Full Spectrum Flow Cytometry with the Cytek Aurora. The video is about 1h 40 min includes both a good discussion of the theory behind the instrument and a hands-on portion demonstrating how to use the instrument. They also offer a second training on the specifics of panel design for full spectrum flow cytometry.
has great videos short videos (~10 minutes each) on their website that demonstrate the basic functions of their cytometers.
Optimized Multicolor Immunofluorescence Panels (OMIPs) are peer-reviewed panels designed for fluorescent assays. They serve as a great starting point for designing a new panel.
has recently released their SpectroLearn™ portal and FSP™ Panel Design Series which takes a dive deep into Full Spectrum Profiling™ Technology (FSP™). This video series contains 6 videos covering the following topics:
Panel Design Step 1: Marker Expression and Co-expression
Panel Design Step 2: Fluorochrome Brightness
Panel Design Step 3: Case Study Analysis
Panel Design Step 4.1: The Importance of Reference Controls
Panel Design Step 4.2: Selecting Appropriate Controls
Panel Design Step 5: Evaluation of Marker Resolution in Multicolor Assays
Resources for Optimizing High Parameter Flow Cytometry Panels
Once your panel design is completed, you will need to physically optimize the protocol at the cytometer. Here is some info on how to optimize high parameter panels for flow cytometry.
This is an excellent protocol walking the user through the panel design process: Panel Design and Optimization for High-Dimensional Immunophenotyping Assays Using Spectral Flow Cytometry – Ferrer-Font - 2020 – Current Protocols – Wiley Online Library.
This is an excellent protocol walking the user through the panel optimization process: Panel Optimization for High‐Dimensional Immunophenotyping Assays Using Full‐Spectrum Flow Cytometry - Ferrer‐Font - 2021 - Current Protocols - Wiley Online Library which includes five crucial steps:
Evaluation of reference controls
Evaluation of unmixing of fully stained samples
Evaluation of marker resolution
Assessment of data quality
The following videos show examples of how to implement the above recommendations.
Maria Jaimes: Best practices for reference controls optimization. Recording
Monica DeLay: How to effectively evaluate data quality and performance. Recording
A feature of the instruments is the ability to extract the autofluorescence. If you are analyzing highly cells, I recommend watching the following brief video Extracting Autofluorescence.
If you are working with very heterogenous samples with multiple signatures, I recommend watching the following videos:
For more advanced users, has a series of presentations on the following topics: