Vybrant™ DyeCycle™ Violet Stain (Invitrogen™)

Vybrant™ DyeCycle™ Violet Stain is a cell permeable DNA dye that can be used for cell cycle analysis and stem cell side population by flow cytometry.

• Precise—accurate cell cycle analysis in living cells

• Safe—low cytotoxicity for cell sorting and additional live cell experiments

• Minimal compensation—easier multiplexing

• Flexible

• Simple, robust staining protocol

View a selection guide for all products related to cell cycle analysis of fixed and live cells in flow cytometry.

Precise

Successful cell cycle analysis requires a dye that is DNA selective and can stain cells in a homogeneous pattern minimizing fluorescence variability. The Vybrant™ DyeCycle™ Violet Stain is an ideal tool for DNA content analysis in living cells since the stain is cell permeable and, after binding double-stranded DNA, emits a fluorescent signal that is proportional to the DNA mass (see figure).

Low Cytotoxicity

Unlike stains that require high concentrations or have chemical structures that are toxic to cells, Vybrant™ DyeCycle™ stains exhibit relatively low cytotoxicity, allowing the possibility of sorting based on the phase of the cell cycle.

Minimal Compensation

Well-suited for the popular violet laser line, the Vybrant™ DyeCycle™ Violet Stain/DNA complex has fluorescence excitation and emission maxima of 369/437 nm, respectively (see figure).

The violet excitation and narrow emission of Vybrant™ DyeCycle™ Violet Stain make it ideal for multiplexing due to the limited spectral overlap with other common dyes (Alexa Fluor™ 488, FITC, and RPE) and fluorescent proteins (Green Fluorescent Protein (GFP) and mCherry) . Vybrant™ DyeCycle™ Violet Stain can also be used with UV excitation, having emission at ∼440 nm.

Flexible

Vybrant™ DyeCycle™ Violet Stain has been shown to not only work for both live cells and fixed cells in cell cycle assays, but to identify stem cell side populations and early progenitors in mammalian hematopoietic tissues (see figure).

Simple, Robust Staining Protocol

For cell analysis, simply prepare flow cytometry tubes each containing 1 mL of cells suspended in complete media at a cell concentration of 1 × 10⁶ cells/mL. To each tube, add 1 μL of Vybrant™ DyeCycle™ Violet Stain and mix well. Final stain concentration is 5 μM. Incubate at 37°C for 30 minutes, protected from light. Keep cells at 37°C until acquisition. Analyze samples without washing or fixing on a flow cytometer using ∼405 nm excitation and ∼440 nm emission.