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Cyto3D Live-Dead Assay Kit

The Cyto3D™ Live-Dead Assay Kit is developed for accurate determination of live/dead nucleated cells by using a dual-fluorescence system. This kit is recommended for viability analysis of cell cultures in 3D or 2D coating or on monolayer.

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Product Description

Acridine orange (AO) and propidium iodide (PI), both nuclear staining (nucleic acid binding) dyes, are used in this kit. AO is permeable to both live and dead cells and stains all nucleated cells to generate green fluorescence. PI only penetrates the membranes of nucleated cells with compromised membranes and stains the dead cells to generate red fluorescence. Due to the quenching, when cells are stained with both AO and PI, all live nucleated cells fluoresce green and all dead nucleated cells fluoresce red (the PI reduces the fluorescence intensity of the AO by fluorescence resonance energy transfer (FRET)). Non-nucleated materials such as red blood cells, platelets and debris do not fluorescence and are ignored fluorescence microscopes.

Dual-Fluorescence Viability, using AO and PI, is the recommended viability analysis method for cell lines, primary cells, and stem cells.

Easy setup and use


Figure 1. Live-dead cell viability images of stem cell spheroids.
Stem cells were static suspension culture in VitroGel STEM (cat# VHM02) for 5 days. 2 µL of Cyto3D reagent was added to each well containing 100 µL cell suspension.  The mixture was incubated at 37 °C for 5-10 min. The cells were then observed under a fluorescent microscope. The images show the Live (green) and Dead (orange) stem cell spheroids cultured in a 3D hydrogel matrix. The live-dead dyes of Cyto3D Live-Dead Assay Kit can successfully penetrate into the big cell spheroids for cell viability analysis.


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