ViaFluor™-SE Cell Proliferation Dyes are fluorescent tracers that diffuse passively into cells and covalently label intracellular proteins, resulting in long term cell labeling. They are initially non-fluorescent, but are converted to amine-reactive fluorescent dyes after hydrolysis by intracellular esterases. The dyes then covalently react with amine groups on intracellular proteins, forming fluorescent conjugates that are retained in the cell, while excess unreacted dye is washed away. Immediately after staining, a single, bright fluorescent population will be detected by flow cytometry.
With each cell division, daughter cells inherit roughly half of the fluorescent label, allowing the number of cell divisions that occur after labeling to be detected by the appearance of successively dimmer fluorescent peaks on a flow cytometry histogram compared to cells analyzed immediately after staining (Figs. 1 and 2).
Cell proliferation dyes can be used to track cell divisions in vivo or in vitro. The staining also can withstand fixation and permeabilization for subsequent immunostaining. ViaFluor™568-SE Cell Proliferation Dye has excitation/emission at 570/606 nm (Figure 3) for detection in the PE-Texas Red® channel with blue laser excitation, or detection in the Texas Red® channel with green laser excitation.
Each kit contains 10 lyophilized dye vials, anhydrous DMSO for preparing stock solutions, and a labeling protocol. One kit can be used to label at least 2×108 cells; the final number of assays that can be performed per kit depends on the dye concentration used (see product protocol under downloads). Biotium also offers blue fluorescent ViaFluor™405-SE and green fluorescent ViaFluor™CFSE Cell Proliferation Kits. [one_half padding=”0 10px 0 0″]
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