StarBright Yellow Dyes

SBY575 is the first of our new StarBright Yellow Dye series. It is a bright 561 nm excitable dye emitting at 579 nm with a narrow excitation and emission profile (Figure 3).

Fig. 3. SBY575. A, Conventional spectra. Excitation by the yellow-green laser (dotted line) with emission shown in the shaded histogram. B, Normalized dye signature. Full emission spectrum showing the emission at all wavelengths. Data were collected on a 5-L Cytek Aurora Flow Cytometry System using SpectroFlo Software.

This results in less spillover when building panels containing 488 nm excitable dyes in conventional flow cytometry.

As it has a unique spectrum and a similarity score of only 0.42, StarBright Yellow 575 Dye can be used with PE in full spectrum flow cytometry.

Although it is not as bright as PE, it is still a bright dye (Figure 4) with the benefit of not being excited to the same extent by the 488 nm laser.

SBY605 is a bright 561 nm excitable dye with a unique and narrow emission that unlike PE-tandems with similar emissions, is not excited by the 488 nm laser (Figure 5).

Fig. 5. SBY605. A, Conventional spectra. Excitation by the yellow-green laser (dotted line) with emission shown in the shaded histogram. B, Normalized dye signature. Full emission spectrum showing the emission at all wavelengths. Data were collected on a 5-L Cytek Aurora Flow Cytometry System using SpectroFlo Software .

Suitable for use in conventional and full spectrum flow cytometry and compatible with other StarBright Dyes without the requirement for a special buffer, it is as bright as PE-Dazzle594 when excited by the 561 nm laser (Figure 6).

SBY605 is not a traditional tandem dye so unlike PE-based tandems can be fixed in PFA or alcohol-based fixatives with no loss of signal or changes to the spectral profile giving you reduced spillover and more flexibility in your experiments.
 

SBY665 is a bright 561 nm laser excitable dye emitting at 670 nm with a unique and narrow emission that, unlike PE-tandems with similar emissions such as PE-Cy5 and PE-A647, is not excited by the 488 nm laser to the same extent, resulting in lower spillover and compensation (Figure 7).

Fig. 7. SBY665. A, Conventional spectra. Excitation by the yellow-green laser (dotted line) with emission shown in the shaded histogram. B, Normalized dye signature. Full emission spectrum showing the emission at all wavelengths. Data were collected on a 5-L Cytek Aurora Flow Cytometry System using SpectroFlo Software .

SBY665 is suitable for use in conventional and full spectrum flow cytometry with a unique spectrum and is compatible with other StarBright Dyes without the requirement for a special buffer. It is brighter than PE-A647, but not as bright as the very bright dye PE-Cy5, when excited by the 561 nm laser (Figure 8).

SBY665 is not a traditional tandem dye so, unlike PE-based tandems, it can be fixed in PFA or alcohol-based fixatives with no loss of signal and minimal changes to the spectral profile giving you reduced spillover and more flexibility in your experiments.
 

SBY720 is a bright 561 nm excitable dye with a unique and narrow emission (Figure 9). It can be used in place of PE-Cy5.5 and PE-A700 in multicolor panels as it is brighter, has reduced 488 nm excitation, and is fixable in PFA and alcohol-based buffers without loss of signal or changes to the spectra (Figure 10). As it has a similarity score of 0.5, it is also suitable to be used with PE-Cy5.5.

Fig. 9. SBY720. A, Conventional spectra. Excitation by the yellow-green laser (dotted line) with emission shown in the shaded histogram. B, Normalized dye signature. Full emission spectrum showing the emission at all wavelengths. Data were collected on a 5-L Cytek Aurora Flow Cytometry System using SpectroFlo Software.

Furthermore, as it is not a traditional tandem dye it is not prone to changes in emission over time due to acceptor and donor uncoupling, giving you reproducible data.

Like all StarBright Dyes, SBY720 can be multiplexed with other StarBright Dyes or other fluorophores without the need for a special buffer and it can be premixed in a panel and stored at 4⁰C for over a month without changes to staining.
 

SBY800 is a bright 561 nm laser excitable dye, which emits at 788 nm with a unique and narrow emission that, unlike PE-Cy7, is not excited by the 488 nm laser to the same extent (Figure 11). Although not as bright as PE-Cy7 (Figure 12), it is still a bright dye that does not bind monocytes in a non-specific manner, unlike CyDye containing tandems.

Fig. 11. SBY800. A, Conventional spectra. Excitation by the yellow-green laser (dotted line) with emission shown in the shaded histogram. B, Normalized dye signature. Full emission spectrum showing the emission at all wavelengths. Data were collected on a 5-L Cytek Aurora Flow Cytometry System using SpectroFlo Software.

SBY800 is not a traditional tandem dye so, unlike PE-based tandems, it can be fixed in PFA or alcohol-based fixatives with minimal loss of signal or changes to the spectral profile, giving you reduced spillover and more flexibility in your experiments.
 

StarBright Yellow Dyes can be used successfully in combination with other StarBright Dyes and traditional fluorophores. Below are comparative data of PE and PE-tandems compared to SBY Dyes in a ten-color panel (Figure 13) . Spreading is reduced in the panel containing SBY Dyes. Additional data featuring StarBright Yellow Dyes and all StarBright Dyes can be found in posters we have presented at conferences.

Fig. 13. Ten-color panel comparing PE and PE-tandem dyes with SBY Dyes.  Human peripheral blood was stained for CD20 (MCA1710F), CD19 (MCA1940SBB580), CD45RA (MCA88SBB675), CD33 (MCA1271SBB810), and CD8 (MCA1226A647). For comparison, additional stains of the following were used: CD4 (MCA1267SBY575 or MCA1267PE), CD3 (MCA463SBY605 or PE-Dazzle594), CD45RO (MCA461SBY665 or PE-Cy5), CD45 (MCA87SBY720 or PE-Cy5.5), and CD14 (MCA1568SBY800 or MCA1568P750). Red boxes highlight the additional spreading seen from PE and PE-tandems. Black arrows show the gating. Data generated on the ZE5 Cell Analyzer.