StarBright Violet 610 Dye
StarBright Violet 670 (SBV670) 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 667 nm, it can be detected using the 670/30 filter on the ZE5 Cell Analyzer or similar filters on other instruments.
Fig. 1. Excitation and emission spectra for SBV670 Dye. 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.
SBV670 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).
Fig. 2. Dye signature of SBV670. Full emission spectrum of StarBright Violet 670 Dye showing the emission at all wavelengths. Data was collected on the Cytek Aurora Flow Cytometry System using SpectroFlo Software.
SBV670 Dye has been designed to be bright with narrow excitation and emission, making it a great choice for inclusion in multicolor panels. It is as bright or brighter than dyes with similar excitation and emission profiles (Figure 3), allowing detection of rare and low antigen density populations. In addition, as it is not a tandem dye, it does not suffer from unwanted excitation by longer wavelength lasers.
Fig. 3. Brightness comparison. A, human peripheral blood was stained with CD4SBV670 (MCA1267SBV670) or CD4BV650 and B, mouse peripheral blood was stained with CD4SBV670 (MCA2691SBV670) or CD4BV650. Both antibodies were titrated prior to use to determine the optimal concentration.
SBV670 Dye 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 4A). StarBright Violet 670 Dye is not a tandem dye so is not susceptible to photobleaching and the staining does not deteriorate with fixation. It also has high lot-to-lot reproducibility and is stable at 4oC with no loss of signal. Furthermore, StarBright Dyes can be used at 4oC or at RT with the optimal staining achieved after at least one hr (Figure 4B), providing the flexibility to fit into any experiment.
Fig. 4. Experimental flexibility. A, human peripheral blood was stained with CD4SBV670 (MCA1267SBV670) in PBS 1% BSA, bovine calf serum containing Staining Buffer (BUF073) or Brilliant Stain Buffer. B, human peripheral blood was stained with CD4 (MCA1267SBV670) at 4oC or RT for 10, 30, and 60 min prior to analysis.
StarBright Violet 670 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.
Fig. 5. Examples of SBV670 Dye staining. Human peripheral blood was stained with A, CD20A488 (MCA1710A488) and CD19SBV670 (MCA1940SBV670); B, CD3PE-Cy5 (MCA463C) and CD27SBV670 (MCA755SBV670); and C, CD45ROSBV440 (MCA461SBV440) and CD45RASBV670 (MCA88SBV670). 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 page.
For more information about fluorophores and immunophenotyping, refer to our flow cytometry resources.
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