ViaFluor® SE Cell Proliferation Kits use amine-reactive dyes to covalently label cells throughout the cell cytoplasm and intracellular compartments. Cell proliferation dyes are commonly used to monitor cell division by flow cytometry. The dyes also can be used to stably label cells to image cell morphology, or to track cell populations in mixed co-culture experiments. Labeling is covalent, so it withstands fixation and permeabilization for subsequent immunostaining. 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. Thus, cell proliferation dyes can be used to track multiple cell divisions of cells grown in culture or injected in vivo after labeling with the ViaFluor® SE dye. Kit Components:
- 10 lyophilized dye vials (full size kits)
- 1 lyophilized dye vial (trial size kits)
- 1 vial of anhydrous DMSO for preparing stock solutions
When used at 1 uM in 1 mL of cells, at a cell density of 1 x 10
6 cells mL, each dye vial can be used for 90-100 labelings; the final number of assays that can be performed per kit depends on the dye concentration used. ViaFluor® kits come with 3 dye options:
- ViaFluor® CFSE (also known as CFDA-SE) can be detected in the FITC channel.
- ViaFluor® 488 SE, an improved alternative to CFSE; detected in the FITC channel.
- ViaFluor® 405 SE for the violet laser; 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.