Viafluor™ SE Cell Proliferation Kits use amine-reactive dyes to covalently label the cell cytoplasm, and can be used to monitor cell division by flow cytometry.
The membrane permeable compound is non-fluorescent until it enters viable cells, where it is hydrolyzed by cytoplasmic esterase enzymes to releases the fluorescent amine-reactive dye. 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.
Kit Components:
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 1.8×108 cells; the final number of assays that can be performed per kit depends on the dye concentration used (see product protocol under downloads).
ViaFluor kits come in 3 dye options:
- ViaFluor™ CFSE (also known as CFDA-SE) can be detected in the FITC channel.
- ViaFluor™ 405-SE can be detected in the Pacific Blue channel.
ViaFluor™ CFSE can be used as a cytoplasmic stain in yeast. However ViaFluor™ 405-SE does not stain well in yeast.
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