Live/dead cell identification

Discrimination between live and dead cells is fundamental to experimentation in cell biology. Clear, fast, and simple labelling is key in both preparative and analytical implementations of flow cytometry and cell culture. Our three live/dead kits use two spectrally distinct and optimized labels to simultaneously and unambiguously mark both live and dead cells.

These kits can be used with suspension cells and adherent cell samples in various fluorescent microscopy applications.

Cell viability

Live/dead assays can be used for assessing cell viability, providing insights into the health and status of cell populations. Advanced dual-dye labeling techniques allow for precise differentiation between live and dead cells, making them ideal for applications such as monitoring cell cultures, evaluating cytotoxicity, and conducting advanced imaging studies.

Yeast viability

Live/dead assays offer a reliable method to distinguish between live and dead yeast cells in various experimental conditions. They provide clear and precise results when used in applications such as fermentation monitoring, stress response studies, and antifungal drug testing.

Bacterial viability

Live/dead assays allow researchers to accurately differentiate between live and dead bacterial cells and can be used in applications such as antibiotic efficacy testing, biofilm analysis, and environmental monitoring.

Principles and advantages of live/dead cell labelling

Our kits make use of the principle that dead cells lose plasma membrane integrity and become permeable. Additionally, esterase activity is significantly reduced in dead cells.

Two of our Live and Dead cell assay kits (ab115347 and ab270789) use calcein-acetoxymethyl ester (Calcein-AM) to identify live cells using the green fluorescence of calcein positively. The esterified form of calcein (Calcein-AM) is non-fluorescent and membrane-permeant. Once it enters cells, intracellular esterases cleave Calcein-AM into calcein, which is both membrane impermeable (trapped inside the live cell) and green fluorescent. In contrast, dead cells do not stain with Calcein-AM due to a lack of intracellular esterase activity and because any fluorescent calcein can exit through the dead cell’s permeant plasma membrane. While researchers often label dead cells only, positively labeling live cells has significant advantages, especially in cases where confounding debris is present, as when establishing primary cultures or analysing disaggregated tissue, or even just demarcating fragments from whole cells in flow cytometry.

These same two kits also incorporate a separate dye to positively label dead cells (ethidium homodimer in ab115347 and 7-AAD in ab270789). Both of these dyes carry multiple positive charges and can only enter cells with compromised membranes (dead cells). They bind tightly to DNA, causing dead cells to fluoresce very brightly in the red portion of the spectrum. Ethidium homodimer is spectrally similar to classical propidium, but with a greater affinity for double-stranded DNA (dsDNA). 7-AAD has a greater Stokes shift and fluoresces at a somewhat longer wavelength than ethidium/propidium, despite similar excitation characteristics, and may be preferred in cases where greater separation from orange-red labels is desired, for example, in a flow cytometry assay that also uses a PE labelled antibody.

Our third Live and Dead Cell Staining Kit (ab65470) has a simpler and more rapid protocol. It uses propidium to label dead cells as described above. However, its differentiating factor is the inclusion of a cell-permeable green fluorescent molecule that efficiently labels all cells, whether live or dead. The result is that dead cells are both red and green (orange), and live cells are green only.

All our Live and Dead Cell staining kits come with clear protocols and ready-to-use formulations that have been pre-balanced to yield optimal staining in both colors.

FAQ

What types of cells can be used with live/dead assays?

Our live/dead assays are versatile and can be used with a variety of cell types, suspension cells, and adherent cell samples. Please contact us (technical@abcam.com) if you have any questions.

Can these assays be quantified?

Yes, the assays are designed to allow quantitative analysis of live and dead cells in fluorescence microscopy applications. Please refer to the protocol booklets for more details.



Our live/dead assays provide reliable solutions for assessing cell viability, offering researchers essential tools to progress their research.