LiFluor™ Animal Cell Viability Kit provides a two-color fluorescence staining on both live and dead cells using two probes that measure two recognized parameters of cell viability - intracellular esterase activity and plasma membrane integrity. The kit is suitable for use with fluorescence microscopes, fluorescence multiwell plate scanners and flow cytometers and other fluorescence detection systems. The assay principles are general and applicable to most eukaryotic cell types, including adherent cells and certain tissues, but not to bacteria or yeast. It is generally faster, less expensive, safer and a more sensitive indicator of cytotoxic events than alternative methods. Live cells are distinguished by the presence of ubiquitous intracellular esterase activity, determined by the enzymatic conversion of the virtually nonfluorescent cell-permeant calcein AM to the intensely fluorescent calcein. The polyanionic dye calcein is well retained within live cells, producing an intense uniform green fluorescence in live cells (Ex/Em ~495 nm/~520 nm). Propidium iodide (PI) enters cells with damaged membranes and undergoes a 30-fold enhancement of fluorescence upon binding to nucleic acids, thereby producing a bright red fluorescence in dead cells (Ex/Em ~528 nm/~617 nm). PI is excluded by the intact plasma membrane of live cells.
Key Features
• Simplicity: Reagents are simultaneously added, no wash steps are required.
• Specificity and reliability: Distinct color for live and dead cell.
• Versatility: Suitable for fluorescence microscope, flow cytometer or microplate reader.
Specifications
1. Platform: Fluorescence Microscopy, Flow Cytometry
2. Detection Method: Fluorescent
3. Ex/Em: Calcein: 494/517; PI: 535/617 nm
Applications
Cell viability assay
Components
1. Calcein AM: 100 µl
2. Propidium Iodide: 100 µll
Storage
Store at -20°C and protect from light.
Case Study
Live and dead Jurkat cells stained with LiFluor™ Animal Cell Viability Kit. Live cells fluoresce a bright green, whereas dead cells fluoresce red.