StarBright Dyes have been designed to be stable with superior brightness, narrow excitation and emission characteristics, and the flexibility to be included in new and existing experiments. Improved resolution of specific cell populations and minimized spillover and spreading allow you to build bigger, better panels with ease. Conjugated to highly cited immunology antibodies, StarBright Dyes are a great addition to Bio-Rad’s range of fluorescent dyes available for ultraviolet, violet, blue, and yellow lasers suitable for use on all cell sorters, cell analyzers, and spectral instruments.
Bright
Narrow excitation and emission
Flexible to work in all buffers
Compatible with all instruments and reagents
Consistent staining
On key immunophenotyping targets
Bio-Rad has launched multiple StarBright Dyes excitable by the ultraviolet, violet, blue, and now yellow/green lasers on a range of human and mouse markers and streptavidin.
At Bio-Rad, we provide a comprehensive range of useful antibody resources to support your research and experiments.
StarBright Dyes are new, proprietary fluorescent nanoparticles conjugated to Bio-Rad's highly validated antibodies. This video demonstrates StarBright Dyes’ exceptional brightness, narrow excitation and emission, and stability with high lot-to-lot reproducibility. You will also discover how easily they can fit into your flow cytometry experiments and how they can be used in immunophenotyping panels to obtain improved results compared to conventional dyes.
Designed to be as bright, or brighter than competitor dyes. The exceptional brightness of StarBright Dyes allows detection of rare and low antigen density populations with excellent separation of positive from negative populations.
Fig. 1. StarBright Dyes are designed to be brighter than competitor dyes. A, examples of CD4 staining for 488, 405, and 355 nm excitable StarBright Dyes compared to competitors. B, stain index of human peripheral blood stained with various 405 nm excitable StarBright, Brilliant Violet and Super Bright Dyes conjugated to CD4. Data generated on the ZE5 Cell Analyzer.
Narrow emission spectra to minimize spillover and reduced excitation by other common laser lines make StarBright Dyes an ideal choice for inclusion in multicolor panels. Use our Spectraviewer to find the right StarBright Dye for your experiment.
Fig. 2. Examples of improved spectral characteristics of StarBright Dyes. Reduced excitation of StarBright Blue 700 Dye, compared with PerCP-Cy5.5 (red circle), and reduced excitation by the 355 nm laser with narrow emission for StarBright Violet 515 Dye, compared to Brilliant Violet 510 (blue circles).
StarBright Dyes have no drop in performance regardless of the staining buffer, and do not require a special staining buffer when multiplexing with each other. However, if special buffers are required due to the presence of multiple polymer dyes in your panel, the flexibility of StarBright Dyes means you will still achieve bright, reliable staining.
Fig. 3. Examples of staining of human peripheral blood in different buffers. A, no change in CD4 staining (MCA1267SBV670) is observed, regardless of the buffer. B, StarBright Dyes, in this case CD4SBV790 (MCA1267SBV790), work well in both Bio-Rad Staining Buffer or Brilliant Staining Buffer with one Brilliant Violet Dye. Data generated on the ZE5 Cell Analyzer.
Designed for multiplexing using the ZE5 Cell Analyzer, StarBright Dyes are suitable for cell analysis or cell sorting on any instrument with the appropriate lasers and filters. StarBright Dyes are compatible with all fluorescently-labeled antibodies and can even be added to premixed cocktails without the need for experimental changes. Furthermore, their unique spectra make them ideal for incorporating into spectral flow cytometry panels with novel dye combinations now possible.
Below are thumbnails of posters which show examples of high parameter panels in conventional and full spectrum flow cytometry. These, along with other useful flow cytometry posters, can be downloaded for your own use.
27-color spectral panel built using StarBright Dyes and a spectral flow cytometer.
Designed with minimum lot-to-lot variability, StarBright Dyes give the same level of staining and population identification regardless of the batch. Unlike some polymer and other dyes, they are not traditional tandem dyes and therefore exhibit less cross-laser excitation, emitted by the donor and acceptors in tandem dyes, due to insufficient FRET. They are resistant to photobleaching, stable without loss of performance over time, and can be fixed in paraformaldehyde or alcohol-based fixatives with no drop in signal or change in their spectral characteristics. Conveniently, they can even be premixed for up to 28 days without loss of performance for use at a later date. To help you get consistently reproducible results, refer to the StarBright Dyes staining protocol.
Fig. 4. StarBright Dyes — consistent, stable, and fixable. StarBright Dyes show consistent results regardless of the lot. A, multiple batches were used to stain peripheral blood mononuclear cells with CD4SBV515 (MCA1267SBV515), and the results overlaid on a histogram. Dotted line is CD4BV510 for comparison. B, CD14SBV610 (MCA1568SBV610) was left at room temperature in the dark or in the light for up to 14 days prior to staining human peripheral blood, and the staining compared to antibody stored correctly in the fridge. A reduction in staining was only seen after 14 days storage at room temperature in the light. C, fixation in Bio-Rad Fixation Buffer or 2% PFA in PBS has no effect on the stain index compared to freshly stained samples. Data generated on the ZE5 Cell Analyzer.
StarBright Dyes are available directly conjugated to antibodies validated for flow cytometry to detect human and mouse targets, and streptavidin for binding to biotinylated antibodies. These antibodies are well-known clones which are highly cited, giving you confidence in your experiments.
Fig. 5. Examples of staining of common immunophenotyping markers using StarBright Dyes. Human peripheral blood was stained for CD3 (MCA463SBUV400), CD19 (MCA1940SBUV605), CD4 (MCA1267SBUV510), CD8 (MCA1226SBV710), CD25 (MCA2127SBV440), and CD127 (HCA145A647) to identify B and T lymphocytes and T helper, T cytotoxic cells, and T regulatory cells within the T cell population. Classical, intermediate, and nonclassical monocytes were identified using CD14 (MCA16568SBV790) and CD16 (MCA2537A700). Further characterization of the T helper cells was obtained from CD45RA (MCA88SBV515) and CD45RO (MCA461SBV610) to identify naïve and memory cells, and the co-stimulation marker expression on T regulatory cells was measured by CD27 (MCA755SBV670) and CD28 (MCA709SBUV575) levels. Data generated on the ZE5 Cell Analyzer.
This course is for anyone who would like to have a detailed understanding about the basics of flow cytometry and experimental best practice. Each month a new online class will be available, and you can join Bio-Rad’s flow cytometry community to get your questions answered.
Mouse Anti-Human CD4 clone RPA-T4 detects the surface glycoprotein human CD4 primarily expressed on T helper lymphocytes, T regulatory lymphocytes, peripheral blood monocytes, and tissue macrophages. Conjugated to all StarBright Dyes, CD4 is ideal for calculating and comparing stain indices between fluorescent dyes.
In this webinar, we will demonstrate how the exceptional brightness of StarBright Dyes along with narrow excitation and emission profiles, their stability, and high lot-to-lot reproducibility can help overcome common challenges when building flow cytometry panels.
It is aimed at anyone who would like to have a detailed understanding of the basics of flow cytometry and stay ahead in all experiments. Led by flow cytometry experts who have diligently compiled all the questions they are regularly asked about the application and have designed this program to be both comprehensive and efficient.
To give you as much flexibility as possible, rather than purchasing pre-planned panels of fluorescent dyes, any sample size of StarBright Dye conjugated CD4 can be purchased individually. A discount is available when buying multiple vials of sample size, increasing when you purchase more.
Mouse Anti-Human CD14 recognizes the human cell surface antigen CD14 which functions as a pattern recognition receptor in innate immunity for a variety of ligands. It is strongly expressed on the surface of monocytes and macrophages but has also been shown to be expressed on the surface of non-myeloid cells.