Fluorophores for the Green (532 nm) and Yellow (561 nm) lasers

Phycoerythrin is a member of the light harvesting phycobiliproteins found in red algae. PE is one of the brightest fluorophores, but is rapidly photobleached, as are tandems of PE, making them unsuitable for fluorescent microscopy applications.

Fig. 1. PE staining. Human peripheral blood was stained with CD4 (MCA1267PE) and CD3 (MCA463A700) after incubation with 10% human serum to detect helper T cells. Data was acquired on the ZE5 Cell Analyzer.

StarBright Yellow 575 Dye is part of the StarBright Dye range, a new range of fluorescent nanoparticles specifically designed for flow cytometry to be bright with narrow excitation and emission profiles. Excitable exclusively by the yellow, 561 nm laser and emitting at 579 nm, this bright fluorophore is the ideal dye to replace PE in panels where the 488 nm laser is being used.

StarBright Yellow 605 Dye is part of the StarBright Dye range. Excitable exclusively by the yellow, 561 nm laser and emitting at 606 nm, this bright fluorophore is the ideal dye to replace PE-DyL594 and PE-Dazzle in panels where the 488 nm laser is being used.

PE-A647 is a tandem dye of PE and Alexa -Fluor 647, exclusive to Bio-Rad. It cannot be used with PE-Cy5 as it has similar excitation and emission characteristics. However, it is a good alternative as it does not exhibit the nonspecific binding to Fc receptors, particularly on monocytes, seen with PE-Cy5.

Fig. 2. PE-Alexa Fluor 647 staining. Murine bone marrow was stained with CD11b (MCA711P647) and GR-1 (MCA2387A488) after incubation with 10% mouse serum to identify monocytes and granulocytes. Data acquired on the ZE5 Cell Analyzer.

StarBright Yellow 665 Dye is part of the StarBright Dye range. Excitable exclusively by the yellow, 561 nm laser and emitting at 670 nm, this bright fluorophore is the ideal dye to replace PE-Cy5 in panels. As it is not a traditional tandem it does not have excitation by the 488 and 640 lasers of the donor and acceptor dyes.

PE-Cy5 is a tandem of PE and Cyanine 5. It cannot be used with PE-A647 or APC due to similar emission spectra. Cy5 in this tandem has been shown to bind nonspecifically to Fc receptors.

PE-Cy5.5 is a tandem of PE and Cyanine 5.5 which has been shown to have less nonspecific Fc receptor binding.

StarBright Yellow 720 Dye is part of the StarBright Dye range. Excitable exclusively by the yellow, 561 nm laser and emitting at 719 nm, this bright fluorophore is the ideal dye to replace PE-Cy5.5 in panels. It has reduced 488 nm laser excitation and unlike Cy-based dyes does not bind to monocytes.

PE-A750 is a tandem dye of PE and Alexa-Fluor 750, exclusive to Bio-Rad. It cannot be used with PE-Cy7 as it has similar excitation and emission characteristics. However, it is a good alternative as it is more stable and not as sensitive to paraformaldehyde fixation as PE-Cy7.

PE-Cy7 is a tandem of PE and cyanine 7. PE-Cy7 is very photosensitive and can degrade with paraformaldehyde fixation. It cannot be used with PE-A750 due to similar emission spectra.

StarBright Yellow 800 Dye is part of the StarBright Dye range. Excitable exclusively by the yellow, 561 nm laser and emitting at 788 nm, this bright fluorophore is the ideal dye to replace PE-Cy7 in panels. It has reduced excitation by the 488 laser and does not bind non-specifically to monocytes, unlike PE-Cy7.