Primary Antibodies for Green and Yellow Lasers

Flow cytometry analysis with a green or yellow laser

With the growing availability of flow cytometers equipped with green (532 nm) and yellow (561 nm) lasers, green light–excitable and yellow light–excitable fluorophores are playing an increasingly critical role in standard multiparametric flow cytometry. The expansive Alexa Fluor dye and eFluor dye portfolios support multicolor immunophenotyping by flow cytometry and includes a variety of dyes compatible with the 532 and 561 nm spectral lines of the green and yellow lasers, respectively.

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Step 1. Search for the marker of interest (e.g., “CD4” or “CD45”).
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Properties of fluorescent dyes for green and yellow lasers

The table below lists the properties of the most commonly used fluorophores for green and yellow lasers.

FluorophoreFluorophore typeExcitation max /Emission max (nm)Relative intensity* (1 = lowest, 5 = highest)Suggested bandpass filter (nm)Uses
Alexa Fluor 532Organic dye532/5543555/20Yellow-fluorescent dye with excitation ideally suited for the 532 nm laser
R-Phycoerythrin (R-PE, PE)Protein496, 565/5755575/26Widely used fluorophore with a high quantum yield that is ideal for detection of low-density antigens
PE-Texas RedTandem496, 565/6153590/40Tandem dye, resonance energy transfer from PE molecule to Texas Red dye
PE-Alexa Fluor 610Tandem496, 565/6283590/40Tandem dye, resonance energy transfer from PE molecule to Alexa Fluor 610 dye
PE-eFluor 610Tandem496, 565/6073610/20Tandem dye, resonance energy transfer from PE molecule to eFluor 610; can be used as a brighter alternative to PE-Texas Red dye
PE-Cyanine 5 (TRI-COLOR)Tandem496, 565/6673670/14Tandem dye, resonance energy transfer from PE molecule to the Cyanine 5 dye
PE-Cyanine 5.5Tandem496, 565/6944695/40Tandem dye, resonance energy transfer from PE molecule to the Cyanine 5.5 dye
PE-Alexa Fluor 700Tandem496, 565/7233695/40Tandem dye, resonance energy transfer from PE molecule to Alexa Fluor 700 dye
PE-Cyanine 7Tandem496, 565/7674780/60Tandem dye, resonance energy transfer from PE molecule to Cyanine 7 dye
* The relative intensity of any fluorophore is dependent upon cell type being studied, reagent used, instrument and instrument configuration. The relative intensities provided are to be used as a guideline; observed intensities may vary.

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