Search Thermo Fisher Scientific
Search Thermo Fisher Scientific
The LIVE/DEAD Viability/Cytotoxicity Assay Kit provides a two-color fluorescence cell viability assay that is based on the simultaneous determination of live and dead neural stem cells (NSCs) with probes that measure two recognized parameters of cell viability: intracellular esterase activity and plasma membrane integrity.
The polyanionic dye calcein AM is well-retained within live cells, producing an intense uniform green fluorescence in live cells (excitation/emission ~495 nm/~515 nm), while ethidium homodimer-1 (EthD-1) enters cells with damaged membranes to produce a bright red fluorescence in dead cells (excitation/emission ~495 nm/~635 nm).
Protocols are provided for fluorescence microscopy or microplate analysis of adherent cells, or flow cytometry analysis of cells in suspension.
Note: Calcein and EthD-1 can be viewed simultaneously with a conventional fluorescein longpass filter. The fluorescence from these dyes may also be observed separately; calcein can be viewed with a standard fluorescein bandpass filter and EthD‑1 can be viewed with filters for propidium iodide or Texas Red dye.
Adherent NSCs may be cultured on sterile glass coverslips or in a multiwell plate.
Allow all the reagents to come to room temperature before proceeding.
Figure 1 - Flow cytometry viability assay using the LIVE/DEAD Viability/Cytotoxicity Kit. A 1:1 mixture of live and ethanol-fixed human B cells was stained with calcein AM and EthD-1 following the protocol provided. Flow cytometry analysis was performed with excitation at 488 nm. The resulting bivariate frequency distribution shows the clear separation of the green fluorescent (530 nm) live cell population from the red fluorescent (585 nm) dead cell population.
LT144 17-Mar-2011