Panel Design Overview
Before designing a panel, you should gather some information:
- A list of markers for your panel
- Expression level of markers on your cells of interest (high/low/unknown)
- A rough gating strategy
- Determine which instrument you are using and which fluorophores are best for that instrument
Then you can begin to assemble your panel:
- Spread out your markers evenly over the lasers (i.e. don’t use 5 violet laser fluorophores for a 6-color panel)
- Assign highly expressed markers to dim fluorophores and markers with low or unknown expression to bright fluorophores
Flow Basics 2.0 is a 5-part training course that covers the practical steps for designing and executing a flow cytometry experiment. The third part covers steps for successful panel design. The full course can be found on the training courses page.
Check out this blog post to learn about common panel design mistakes that lead to bad data.
Resources for CAT Facility Instruments
Attune NxT 4-14
Fortessa 4-15 HTS
External Websites with Panel Design Tools
Registration may be required
Our instruments are listed on Fluorofinder. The website will guide you through the panel design steps: enter your marker, target species, antigen density, etc.
The result is a list of available antibodies paired with fluorophores from various manufacturers.
BenchSci is an AI-Assisted antibody selection tool. Simply enter the desired antibody and you get a list of figures and publications related to that antibody.
CiteAb provides a way to choose reagents by reviewing the scientific literature.
Antibodypedia scores antibodies to guide researchers to choose an appropriate antibody for a particular application.
Optimized Multicolor Immunofluorescence Panels (OMIPs)
Optimized Multicolor Immunofluorescence Panel (OMIP) is 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.
OMIPs are aimed: (1) to alleviate the development time for researchers in need of the same or highly similar panels, (2) to provide a starting point for the creation of novel OMIPs, and (3) to give the developers of the panels credit via citation or the publication.
Useful information/web links
- Antibody Fixation Consideration (Thermofisher)
- Antibody Fixation Consideration (BioLegend)
- Phospho-Buffer Consideration (ThermoFisher_
- PhosphoFlow Buffer compatibility (BD Biosciences)
- Enzyme explorer for tissue dissociation (Millipore Sigma)
- Essential Markers (BioLegend)
ChUG empower researchers with knowledge of cytometry tools and applications needed to generate impactful discoveries. ChUG is a scientific educational group based in the Greater Chicagoland area that aims to connect researchers in the community to facilitate the transfer of knowledge in areas of cytometry technology and methodology.
Established in 1989, the list has been continuously available for free access to all scientists. It is a vibrant community of flow cytometry users and a great place to learn about the field and get help from experts around the world. The current membership is approximately 4500 scientists. Subscribe here.