StarBright Violet 710 Dye
StarBright Violet 710 (SBV710) Dye is the latest addition to the StarBright Dye range of fluorescent nanoparticles specifically designed for flow cytometry.
Excitable by the violet 405 nm laser and emitting at 713 nm, it can be detected using the 720/60 filter on the ZE5 Cell Analyzer or similar filters on other instruments.
Fig. 1. Excitation and emission spectra for SBV710. The dye is excited maximally by the violet laser (dotted line) with minimal excitation by other lasers. The emission profile is narrow to reduce spillover into neighboring filters.
StarBright Violet 710 Dye is also compatible with spectral flow cytometry and has a unique profile, allowing incorporation into new and existing flow cytometry panels. The full spectral profile is shown below (Figure 2). This spectra is sufficiently different from Brilliant Violet 711 (BV711), with a similarity index of 0.96, for the two dyes to be used simultaneously in multiplex panels.
Fig. 2. Dye signature of SBV710. The full emission spectrum of StarBright Violet 710 Dye showing the emission at all wavelengths. Data was collected on the Cytek Aurora Flow Cytometry System using SpectroFlo Software.
SBV710 Dye has been designed to be bright with narrow excitation and emission, making it a great choice for inclusion in multicolor panels. It is up to 3x brighter than dyes with similar excitation and emission profiles (Figure 3A), allowing detection of rare and low antigen density populations. In addition, as it is not a traditional tandem dye, it suffers less from unwanted excitation by longer wavelength lasers and has little or no signal from a donor dye (Figure 3B).
Fig. 3. Brightness comparison and spectral properties. A, human peripheral blood was stained with CD4SBV710 (MCA1267SBV710) (red) or CD4BV711 (blue). Both antibodies were titrated prior to use to determine the optimal concentration. SBV710 stain index was 3x that observed for BV711. B, the spectral properties of SBV710 are improved compared with other polymer dyes with similar maximal emission, due to no signal from a donor dye, as can be seen for the 420/10 channel from the 405 nm laser.
Fig. 4. Experimental flexibility. A, human peripheral blood was stained with CD4SBV710 (MCA1267SBV710) in Bio-Rad Staining Buffer containing bovine calf serum (BUF073), Brilliant Stain Buffer, or in PBS 1% BSA.
SBV710 has been tested in most common staining buffers and special staining buffers for polymer dyes, with no drop in performance. No special staining buffer is required, so it can be used in new or existing panels with no change to your experimental protocol (Figure 4).
StarBright Violet 710 Dye is very stable when stored at 4oC, it is not susceptible to photobleaching, and the staining does not deteriorate with fixation. It has high lot-to-lot reproducibility so you can expect the same high quality staining regardless of the lot purchased.
StarBright Violet 710 Dye is compatible with all common fluorophores such as organic fluorophores, protein fluorophores, polymer dyes, and other StarBright Dyes in multicolor panels, without the requirement for a special staining protocol or a special staining buffer. Examples of staining with other dyes, including StarBright Dyes can be seen in Figure 5. If you are using multiple dyes that require a special buffer, we recommend using that buffer as StarBright Dye performance will not be affected.
Fig. 5. Examples of SBV710 Dye staining. Human peripheral blood was stained with A, CD3SBV440 (MCA463SBV440) and CD4SBV710 (MCA1267SBV710); B, CD3BV421 and CD4SBV710 (MCA1267SBV710); and C, CD3A488 (MCA463A488) and CD69SBV710 (MCA2806SBV710). All staining was performed at 4oC in PBS containing 1% BSA after blocking with 10% human serum. Cells were analyzed on the ZE5 Cell Analyzer.
For more information on how the StarBright range can fit into your flow cytometry experiments, please take a look at our StarBright Dyes range.
For more information on fluorophores and immunophenotyping, refer to our cytometry resources.
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