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Part of the new StarBright Dye range, StarBright Violet 610 (SBV610) Dye is a fluorescent nanoparticle. Excitable by the violet 405 nm laser and emitting at 607 nm, it can be detected using the 615/24 filter on the ZE5 Cell Analyzer or similar filters on other instruments.
Fig. 1. Excitation and emission spectra for SBV610. 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 610 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 SBV610. The full emission spectrum of StarBright Violet 610 Dye showing the emission at all wavelengths. Data was collected on the Cytek Aurora Flow Cytometry System using SpectroFlo Software.
Fig. 3. Brightness comparison. Human peripheral blood was stained with CD4 conjugated to StarBright Violet 610 (MCA1267SBV610), or Brilliant Violet 605. Both antibodies were titrated prior to use to determine the optimal concentration.
SBV610 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.
SBV610 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 SBV610 can be used in new or existing panels with no change to your experimental protocol (Figure 4A). StarBright Violet 610 Dye is not a tandem dye so is not susceptible to photobleaching and the staining does not deteriorate with fixation (Figure 4B). Furthermore, SBV610 has high lot-to-lot reproducibility and is stable at 4oC with no loss of signal.
Fig. 4. Experimental flexibility. A, human peripheral blood was stained with CD4SBV610 (MCA1267SBV610) in PBS 1% BSA, bovine calf serum containing Staining Buffer (BUF073) or Brilliant Stain Buffer. B, human peripheral blood was stained with CD14 (MCA1568SBV610) and analyzed fresh, fixed in Fixation Buffer (BUF071) or fixed in a 2% paraformaldehyde solution prepared in-house. The staining was consistent regardless of buffer or fixative.
StarBright Violet 610 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. Protocols have been tested up to one hr at 4oC or RT with no significant alteration in the staining observed. Examples of staining with other dyes, including StarBright Dyes can be seen in Figure 5.
Fig. 5. Examples of StarBright Violet 610 Dye staining. Human peripheral blood was stained with A, CD19 StarBright Blue 700 (MCA1940SBB700) and HLA DP DQ DR StarBright Violet 610 (MCA477SBV610) or B, CD3A647 (MCA463A647) and CD25 StarBright Violet 610 (MCA2127SBV610) after stimulation with PHA for 3 days. 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.
Click on the links below to find out more in-depth information on each topic and view our popular flow cytometry basics guide.