Useful Links
Spectral Viewers
- BD Spectrum Viewer
- Bio-Rad Spectra Viewer
- BioLegend Spectra Analyzer
- Chroma
- FluoroFinder Spectra Viewer
- Omega Optical Spectral Overlap Guide
- ThermoFisher Fluorescence Spectrum Viewer
Antigen Density
How does a Flow Cytometer Work?
- Flow Cytometry Animation
- Flow Cytometry Basics (with a clear explanation of what compensation is)
- Light Scatter Tutorial from CytometryMan
Panel Building Tools
- Bio-Rad Panel Builder: https://www.bio-rad-antibodies.com/flow-panel-builder.html
- BD Horizon Guided Panel Solution (GPS)
- FluoroFinder Panel Builder with UNC configurations. Video Tutorial.
- Optimized Multicolor Immunofluorescence Panels via FluoroFinder (OMIPs – peer-reviewed panels designed for fluorescent assays)
Multicolor Flow Cytometry
- Fluorophore Brightness list from Biolegend – does not include Brilliant UV or Brilliant Blue dyes. Check BD for those.
- The Daily Dongle blog from FlowJo covers tips on Compensation, software updates and links to free webinars.
- Fundamentals of Flow Cytometry (50 minutes)
- Compensation Basics from BD
- Multicolor panel design basics including fluorophore selection and spill over consideration from BD
- BD Horizon Tour videos on precision multi-color panel design, spill-over, compensation, receptor density, and best practices. A fairly non-invasive registration process is required to view the videos, but worth it.
- Basic Flow Cytometry Definitions from Biorad
- Fluorophore combinations to avoid due to increased spreading error. This new version from BD includes the Brilliant Violet and Brilliant UV dyes. Note that the configuration is for the LSRFortessa X20, which is a little different from our core instruments, but the major concepts remain.
- See what the Biexponential Data Spread looks like.
- The impact of adjusting PMT voltages on spillover and compensation – from Sydney Flow Cytometry
- Antibody Fixation considerations from eBio
- Life Technologies School of Fluorescence
- Fuorescent Proteins Table
- Another Fluorescent Protein Table for Flow Cytometry
- Introduction to Fluorescent Proteins (Nikon Microscopy)
- Molecular Probes table of Fluorescent Calcium indicators
- BD Multicolor Fluorochrome Guide
- Excyte Blog on Statistics and Data Analysis
- Excyte Blog on Viability Dyes
- Excyte Blog on Phospho-Flow staining and 4 considerations for Assessing Phosphorylation using flow
- How many cells do I need to collect? ThermoFisher guide to calculating how many cells for a given frequency and desired CV.
- Protocols on cell preparation, staining, cell cycle from Biorad
- Cell Cycle Analysis from Miltenyi Biotec
- 10 Tips and Tricks for the design of multi-color panels – Checklist from Biorad
- Autofluorescence: Causes and Cures
- University of Chicago Flow Cytometry Lab (Ryan Duggan’s Blog)
- FlowMetric – white papers and protocols (clinically oriented)
- BenchSci – free antibody search platform designed to help scientists find antibodies from publications by specific experimental contexts. A.I driven, decodes published figures to help scientists find antibody usage data.
Tutorials (some require a subscription or membership)
- Work-Flow.tech provides e-Learning courses in flow cytometry. Its flagship course, Flow Cytometry Essentials, is a streamlined and user-friendly educational program that demystifies common concepts and serves as a prerequisite to instrument training. Login/subscription required.
- Bio-Rad Flow Cytometry Guide: Bio-rad
- Basic Introduction to Flow Cytometry
- CYTO University – free to ISAC members. Non members can pay a small fee to view specific content.
- Basics of Flow Cytometry (ThermoFisher)
- Flow Cytometry Web-Based training (Becton Dickinson)
- Tips for Good Flow Cytometry Experiments (Chromocyte)
- Fluorescence Tutorials (Life Technologies)
- Webinar on Apoptosis, Cell Signaling, Stem Cells, T-Cell Immunology, Bead-Based Immunoassays (Becton Dickinson)
- FlowJo Training (Tutorials)
- Michael Ormond’s online course
- Paul Robinson’s Purdue Class
- Other lectures on the Purdue Cyto site
Microvesicles and exosomes
- Exosomes by Miltenyi
- Microvescicles and viruses on the ImageStreamX (registrations required)
RNA Detection by Flow Cytometry:
- PrimeFlow Brochure November 2014
- PrimeFlow White Paper November 2014
- Nature Communications Paper (Porichis et al 2014, 5:5641)
- FireFly – Abcam – Firefly is a new platform for multiplexing the measurement of up to 68 microRNAs in a high throughput format. The technology can work directly from either small volumes of biofluids or very low amounts of total RNA.
Reference Publications
- Current Protocols in Cytometry (we have a subscription – let us know if you need any methods)
- Interpreting Flow Cytometry Data: A Guide for the Perplexed (Herzenberg et al. 2006 Nature Immunology 7:681)
- OMPIS (Optimized Multicolor Immunofluorescence Panel) – a special peer-reviewed Cytometry Part A publication type that reports on newly designed and optimized multicolor panels for flow cytometry, fluorescence microscopy, image cytometry, and other polychromatic fluorescence-based methods.
Vendors Websites
- Amnis
- BD Biosciences
- Beckman Coulter
- BioLegend
- Biorad
- Cytek Development
- eBiosciences
- Intellicyt
- Invitrogen
- Miltenyi
- Novus Biologicals – contains query tool that includes UNC cytometers. Has links to basics of flow and protocols.
- Phoenix Flow
- R&D Systems
- Spherotech
- StemCell Technologies
- Stratedigm – Flow Cytometry Systems
Software
- Software List (Chromocyte)
- FlowJo
- Verity Software House (ModFit, WinList, GemStone)
- DeNovo Software/FCS Express
Freeware
Cytometry Groups
- International Society for Analytical Cytology (ISAC)
- Purdue Flow Cytometry (current flow issues and concerns by users over the globe)
BD Researcher’s Digest