Bio-Rad has both traditional fluorophores and new StarBright Dyes that are excitable by the violet (405 nm) laser suitable for conventional flow cytometry and spectral flow cytometry. Table 1 below shows the excitation and emission maxima, the relative brightness, and ZE5 Cell Analyzer filter which allows detection. Other instruments may have different filters, please check compatibility on your instrument before you start your experiment.
Table 1. 405 nm excitable dyes for flow cytometry.
Fluorophore |
Ex max |
Em max |
Brightness |
ZE5 Filter |
Similar |
---|---|---|---|---|---|
StarBright Violet 440 |
383 |
436 |
5 |
460/22 |
Pacific Blue |
Pacific Blue |
410 |
455 |
1 |
460/22 |
SBV440 |
StarBright Violet 475 |
405 |
479 |
4 |
460/22 |
BV480 |
StarBright Violet 515 |
402 |
516 |
4 |
525/50 |
BV510 |
Amethyst Orange |
405 |
540 |
1 |
525/50 |
Pacific Orange |
StarBright Violet 570 |
404 |
571 |
4 |
- |
BV570 |
StarBright Violet 610 |
403 |
607 |
4 |
615/24 |
BV605, SB600 |
StarBright Violet 670 |
401 |
667 |
5 |
670/30 |
BV650, SB645 |
StarBright Violet 710 |
402 |
713 |
5 |
720/50 |
BV711, SB702 |
StarBright Violet 760 |
403 |
754 |
5 |
750LP |
BV750 |
StarBright Violet 790 |
402 |
782 |
4 |
750LP |
BV786, SB780 |
Pacific Blue is a blue, fluorescent, coumarin based dye, excitable by the 405 nm laser. It emits at 455 nm and is best used for markers that are highly expressed due to its dim nature. This fluorophore is not compatible with StarBright Violet 440 Dye due to similar emission spectra and spillover characteristics.
Fig. 1. Pacific Blue staining. Mouse bone marrow was stained for CD11b (MCA711PB) and GR-1 (MCA2387A700) after blocking with Mouse Seroblock (BUF041A) to detect monocytes and granulocytes. Data acquired on the ZE5 Cell Analyzer.
StarBright Violet 440 Dye
StarBright Violet 440 Dye is part of the StarBright Dyes range, a new range of fluorescent nanoparticles specifically designed for flow cytometry to be bright with exact excitation and emission profiles. Excitable by the violet, 405 nm laser and emitting at 436 nm, this bright fluorophore can replace dimmer fluorophores such as Pacific Blue in your flow cytometry panels and is an excellent alternative to Super Bright 436 and Brilliant Violet 421.
StarBright Violet 475
StarBright Violet 475 (SBV475) Dye is part of the range of StarBright Dyes, new fluorescent nanoparticles specifically developed for flow cytometry. Excitable by the violet, 405 nm laser and emitting at 479 nm, this bright dye is a suitable alternative for tandem based polymer dyes with similar emission, such as Brilliant Violet 480, in your multicolor panels.
StarBright Violet 515 Dye
StarBright Violet 515 Dye is part of the StarBright Dyes range, a new range of bright, fluorescent, nanoparticles specially developed for flow cytometry. This bright dye is excitable by the violet, 405 nm laser and emits at 516 nm with reduced spillover into neighboring channels compared to other dyes with similar properties.
Amethyst Orange is a violet laser excitable fluorophore that emits at 540 nm. It is optimally used for high density antigens, or when low level staining is required as it is one of the dimmest fluorophores. This dye gives you increased options for the violet laser when building multicolor panels.
Fig. 2. Amethyst Orange staining. Human peripheral blood was stained for CD4 (MCA1267AMO) and CD3 (MCA463A488) after blocking with 10% human serum. Data acquired on the ZE5 Cell Analyzer.
StarBright Violet 570 Dye
StarBright Violet 570 Dye is part of the StarBright Dyes, a new range of fluorescent nanoparticles specifically developed for flow cytometry. Excitable by the violet 405 nm laser and emitting at 570 nm, this bright fluorophore can replace the polymer dye Brilliant Violet 570. StarBright Dyes are not traditional tandems and have more efficient FRET, resulting in less signal from the acceptor and donor dyes by themselves.
StarBright Violet 610 Dye
StarBright Violet 610 Dye is part of the StarBright Dyes range, a new range of fluorescent nanoparticles specially developed for flow cytometry. Excitable by the violet, 405 nm laser and emitting at 606 nm, this bright fluorophore can replace the polymer dyes Brilliant Violet 605 and Super Bright 600. As it is not a tandem dye, it exhibits less excitation by multiple lasers and is not sensitive to degradation.
StarBright Violet 670 Dye
StarBright Violet 670 Dye is part of the StarBright Dyes, a new range of fluorescent nanoparticles specially developed for flow cytometry. Excitable by the violet, 405 nm laser and emitting at 667 nm, this bright fluorophore is a suitable alternative for tandem based polymer dyes with similar emission, such as Brilliant Violet 650 and Super Bright 645, in your multicolor panels.
StarBright Violet 710 Dye
StarBright Violet 710 Dye is part of the new range of StarBright Dyes, a new range of fluorescent nanoparticles specially developed for flow cytometry. Excitable by the violet, 405 nm laser and emitting at 713 nm, this bright fluorophore is a suitable alternative for tandem based polymer dyes with similar emission, such as Brilliant Violet 711 and Super Bright 702, in your multicolor panels.
StarBright Violet 760 (SBV760) Dye is part of the range of StarBright Dyes, new fluorescent nanoparticles specifically developed for flow cytometry. Excitable by the violet, 405 nm laser and emitting at 754 nm, this fluorescent dye is a brighter alternative for tandem based polymer dyes with similar emission, such as Brilliant Violet 750.
Starbright Violet 790 Dye
StarBright Violet 790 (SBV790) Dye is part of the range of StarBright Dyes, new fluorescent nanoparticles specifically developed for flow cytometry. Excitable by the violet, 405 nm laser and emitting at 782 nm, this bright fluorophore is a suitable alternative for tandem based polymer dyes with similar emission, such as Brilliant Violet 785, Super Bright 780, and SuperNova v786 in your multicolor panels.
Bio-Rad stocks a selection of different fluorophores that are usable with a range of lasers. Details of which fluorophores are best suited to which lasers can be found on our Fluorophores for Flow Cytometry page.
For more information about fluorophores and immunophenotyping, refer to our flow cytometry resources below.