We offer over a dozen web tools to educate researchers on fluorophores, build the ideal multicolor panel, and choose the right reagents for flow cytometry applications. For additional inquiries, contact our Technical Services team

Conventional Spectra Analyzer

“Conventional/traditional” flow cytometers use a series of mirrors and filters in order to capture the emission wavelengths from excited fluorophores. Fluors that have emission profiles that fall into the same filter cannot be separated or identified from one another.

 

Our spectra analyzer can display the excitation and emission spectra for dozens of fluorophores. This allows you to compare the excitation and emission profiles of fluors, custom filters, and laser lines to see what best fits your cytometer.

 

View the Conventional Spectra Analyzer >

Spectral Cytometry Analyzer

Spectral cytometers capture the full spectrophotometric profile of the fluorophores across all lasers. The fluors’ profiles are captured in 10-30 nm segments across the emission range to accurately unmix the fluorophores. This allows fluors that normally cannot be unmixed on a conventional cytometer (e.g. Pacific Blue™ and Brilliant Violet 421™) to be used together on an instrument like Cytek’s Aurora.

 

As such, this web tool is dedicated to displaying the emission spectra of fluorophores based on Cytek’s Aurora spectral cytometers.

 

View the Spectral Cytometry Analyzer >


A comparison of channels that might typically be used for the violet laser on a spectral cytometer (top) compared to filters on a conventional cytometer (bottom). The added “side-by-side” channels on a spectral cytometer allow spectrally similar fluors to be unmixed depending on how their emission profiles look in other channels (including the ones available on additional lasers). On a conventional cytometer, a fluor’s main ‘peak’ or emission is mainly captured in one filter set, making it harder to distinguish spectrally similar fluors.

Conventional Spectra Analyzer

“Conventional/traditional” flow cytometers use a series of mirrors and filters in order to capture the emission wavelengths from excited fluorophores. Fluors that have emission profiles that fall into the same filter cannot be separated or identified from one another.

 

Our spectra analyzer can display the excitation and emission spectra for dozens of fluorophores. This allows you to compare the excitation and emission profiles of fluors, custom filters, and laser lines to see what best fits your cytometer.

 

View the Conventional Spectra Analyzer >

Spectral Cytometry Analyzer

Spectral cytometers capture the full spectrophotometric profile of the fluorophores across all lasers. The fluors’ profiles are captured in 10-30 nm segments across the emission range to accurately unmix the fluorophores. This allows fluors that normally cannot be unmixed on a conventional cytometer (e.g. Pacific Blue™ and Brilliant Violet 421™) to be used together on an instrument like Cytek’s Aurora.

 

As such, this web tool is dedicated to displaying the emission spectra of fluorophores based on Cytek’s Aurora spectral cytometers.

 

View the Spectral Cytometry Analyzer >


A comparison of channels that might typically be used for the violet laser on a spectral cytometer (top) compared to filters on a conventional cytometer (bottom). The added “side-by-side” channels on a spectral cytometer allow spectrally similar fluors to be unmixed depending on how their emission profiles look in other channels (including the ones available on additional lasers). On a conventional cytometer, a fluor’s main ‘peak’ or emission is mainly captured in one filter set, making it harder to distinguish spectrally similar fluors.

BioLegend Panel Selector

BioLegend's Multicolor Panel Selector is designed to help you find the right products for your multicolor flow cytometry experiments. The Panel Selector instantly shows all available BioLegend antibodies for targets of interest and provides easy add-to-cart options for those ready to purchase.

 

View the BioLegend Panel Selector >

FluoroFinder Panel Builder

FluoroFinder’s web tool adds the ability to select your institution’s cytometer configuration to predetermine the fluorophores that can be used in your panels. You can also indicate the expected antigen density on your targets to help balance the fluor brightness selected.

 

View the FluoroFinder Panel Builder >

Gating Strategy

Use our web tool to build your gating strategy and plan complex multicolor flow cytometry experiments. Save your strategy to reference later or share with collaborators to help build your panel.

 

View the gating strategy tool >

Multicolor Staining Guide

With wide varieties of antibodies, fluorophores, and instruments available for multicolor flow cytometry, optimizing your multicolor experiments can be daunting. Use our Multicolor Staining Guide to assist you in developing and optimizing your flow cytometry experiments.

 

View the multicolor staining guide >

Fluorophore Families

Diverse families of fluorophores allow us to continue to expand the limits of flow cytometry. Learn more about our fluors and the unique advantages offered by each, including Spark Dyes, Fire Dyes, KIRAVIA multimers, and Brilliant Violet™ polymers.

 

Learn about fluorophores >

Brightness Index

This convenient Fluorophore Brightness Index lists the relative brightness of dyes offered by BioLegend. This is particularly helpful for the balancing of brightness with antigen expression levels in a panel.

 

View the Brightness Index >

Fluorophore Equivalents

With companies each having their own naming system for their fluorophores, it can be difficult to keep track of them all. This tool will help you find BioLegend's equivalent fluorophores compared to other commercially available fluorophores for flow cytometry.

 

View fluorophore equivalents >

Tandem Dyes

Our tandem dye page provides useful information on the background of tandem dyes, important notes, and how best to use them in flow cytometry applications. We also provide data on lot-to-lot consistency and links to products and resources.

 

Learn about tandem dyes >

Spectral Cytometry

Spectral cytometry is quickly allowing researchers to create larger multicolor panels with its unique technology. Learn the basics of spectral flow cytometry and how BioLegend is providing unique fluorophores to fill spectral gaps previously unused in conventional flow.

 

Learn about spectral cytometry >

Controls

In flow cytometry, having the right controls is just as important as staining your samples. From compensation beads to Fc blocking, our webpage guides you through multiple controls and the situations they're appropriate for. Learn what makes your results real and not artifacts.

 

Learn about flow cytometry controls >

Technical Protocols

Every good experiment begins by having the right protocol set in place. Browse through our flow cytometry protocols, including surface marker staining and intracellular staining for cytokines and nuclear transcription factors.

 

View our protocols >

Instrument Guide

The Flow Cytometry Instrument Guide provides a general overview of which fluorophores can be used on commercially available instruments. Select and compare a wide variety of instruments, including both cell sorters and analyzers.

 

View the Instrument Guide >

Antibody Cross Reactivity

Beyond human and mouse models, researchers also use several other animal models, including non-human primates. This chart, which is based on third-party resources, provides information on whether BioLegend clones have been shown to react to other animal species.

 

View the Antibody Cross Reactivity Tool >

Fixation

Fixation helps to lock proteins in place on cells you plan to analyze. Because this can alter epitopes, it can create problems if samples are fixed prior to staining with antibodies. This webpage provides in-house testing data for several antibody clones, showing staining data on both unfixed cells and cells fixed with 4% paraformaldehyde.

 

View the Fixation Webpage >

Troubleshooting

Our Flow Cytometry Troubleshooting Guide is designed to help plan or optimize your flow cytometry experiments. By covering common topics like low signal, high background, and autofluorescence, you can be prepared for the factors that could impact your experimental results.

 

Learn about troubleshooting >

Contact Technical Services

Our team of scientific experts are ready to help answer any inquiries you may have about our reagents. In addition, they can help you optimize your workflow, aid in troubleshooting, or design multicolor flow cytometry panels.

 

Connect with us >

Autogate Software

This software was built by the Herzenberg Lab at Stanford University. Freely available to non-commercial users, this software provides automation that researchers can control. Researchers can review cell population clusters in their samples, as well as automate all compensation and gating for data sets.

 

View the software >

Flow Cytometry Antibody Conjugates

BioLegend offers an extensive selection of antibody conjugates for flow cytometry. View them all with these handy reference charts to visualize fluorophore conjugate availability for each antibody clone.

 

View flow cytometry conjugates >

Flow Cytometry Buffers

Buffers help create the proper environment for antibody staining applications. View our available buffers for surface and intracellular staining flow cytometry applications.

 

View flow cytometry buffers >

Live/Dead Indicators

It is critical to understand the degree of cell death in any flow cytometry assay and exclude those cells from the analysis. Learn more about our tools for live/dead cell discrimination and the differences between DNA-binding dyes like 7-AAD, and Zombie Dyes, which bind to primary amines on proteins.

 

View live/dead dyes >

Custom Reagents

Our custom solutions team of scientists are ready to help with your specialized projects, whether that means providing bulk sizing of your favorite reagents or creating custom antibody conjugations to fluorophores you can’t find anywhere else. Customize your products to fit your research’s needs.

 

Customize your reagents >

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