Cell Health

Our best assays for measuring cell viability and mitochondrial function

Cell Health

An essential step in early stage, translational research is measuring cytotoxicity. ION’s product portfolio is designed to help you easily capture important parameters of cell health, whether your preferred instrument is a plate reader, fluorescence microscope, or flow cytometer, using a collection of criteria that are indicative metrics of cell viability or death. These criteria include intracellular esterase activity as a marker of cell viability, plasma membrane integrity as a marker of cell death, and mitochondrial membrane potential as an early-stage marker of apoptosis. When used in combination, these assays can provide an informative snapshot of cell health.

 

Header for cell health assays
Fluorescence images of mitochondrial membrane potential in healthy (orange color) and compromised (green) cells.

Intracellular esterase activity

Intracellular enzyme activity and dye retention is a hallmark of viable cells. During cell death, increased plasma membrane permeability causes enzyme leakage and a reduction in activity. Dyes that measure esterase activity are modified with ester-moieties, which help them passively diffuse through cell membranes and render the dyes non-fluorescent. Once inside the cell, ubiquitous esterases present in the cytosol cleave ester-moieties, activating dye fluorescence and trapping the dye within the cell. ION sells Calcein AM in multiple formats (dry or in a prepared solution) and in combination with Ethidium Homodimer I as part of ION Vital – Viability so you can simultaneously detect dead cells.

 

(Ex/Em 495 nm/515 nm)Calcein AM is a membrane-permeant, non-fluorescent form of calcein that is converted to green fluorescent calcein in viable cells, resulting in uniform cytosolic fluorescence. Calcein is well retained within the cytosol of most healthy cells with intact cell membranes.

This kit combines Ethidium Homodimer I and Calcein AM to provide a two-color, fluorescence-based assay to discriminate between live and dead cells.

(Ex/Em 490 nm/535 nm)BCECF AM is the most popular green fluorescent, intracellular pH indicator, however it can also be used as a marker of intracellular esterase activity. For this application, BCECF should be used in non-ratiometric mode using standard FITC excitation and emission settings.

Plasma membrane integrity

A loss of plasma membrane integrity is an indicator of cell death. Nucleic acid-binding, cell impermeant dyes, like Ethidium Homodimer I (EthD-I), can be used to label the nuclei of cells with compromised plasma membranes. These dyes are excluded from intact, healthy cells and exhibit minimal fluorescence in the absence of nucleic acids. Once the dye crosses the cell membrane and binds to nucleic acids (DNA) inside the cell, it displays a significant increase in fluorescence. ION sells EthD-I in multiple formats (dry or in a prepared solution) and in combination with Calcein AM as part of ION Vital – Viability so you can simultaneously detect viable cells.

 

(Ex/Em: 528 nm/617 nm) – EthD-I is a membrane-impermeable, high-affinity, nucleic acid stain that selectively labels dead cells with compromised plasma membranes.

This kit combines Ethidium Homodimer I and Calcein AM to provide a two-color, fluorescence-based assay to discriminate between live and dead cells. 

Mitochondrial membrane potential

Mitochondrial membrane potential is one of the driving forces behind mitochondrial ATP synthesis, and is necessary for maintaining cell homeostasis. During early stages of apoptosis, mitochondrial function and membrane integrity are disrupted by the activation of apoptotic proteins and subsequent release of cytochrome C, mitochondrial Ca2+ stores, and other components. Membrane potential sensitive dyes, such as JC-1 and JC-10, can be used to monitor the loss of mitochondrial membrane polarization observed during apoptosis. These dyes compartmentalize in healthy mitochondria, leading to the formation of aggregates that exhibit orange fluorescence. When mitochondrial membrane potential is lost during apoptosis, these dyes no longer accumulate and return to green fluorescent monomers. ION sells JC-10 individually and as part of our optimized ION Vital Assay line of products

 

JC-10 →

(Ex/Em: 490,540 nm/525, 590nm) JC-10 is a mitochondrial membrane potential probe. It possesses superior aqueous solubility compared to its better known analogue, JC-1. At low concentrations, JC-10 is monomeric and emits a green fluorescence. JC-10 accumulates in healthy mitochondria, forming J-aggregates that exhibit an orange fluorescence. Its potentiometric behavior allows for ratiometric analysis of mitochondrial membrane potential, where a shift from orange to green fluorescence is indicative of compromised mitochondria.

MitoVolt is a mitochondrial membrane potential assay kit that uses JC-10, a potentiometric dye that is more soluble than JC-1. It is an ideal solution for detecting changes in mitochondrial membrane potential due to cell apoptosis or other stress-inducing phenomena, and is compatible with fluorescence microscopy, flow cytometry, and plate reader applications.

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Calcein AM product packaging with cell health sticker, purple background
Calcein AM is a membrane-permeant, non-fluorescent indicator that is converted to green fluorescent calcein by esterase activity in viable cells.
$109.00-$165.00
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Ethidium homodimer product packaging, purple background
  Red fluorescent, dead cell stain  
$165.00-$198.00
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Two-color, fluorescence-based assay to discriminate between live and dead cells
$242.00
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JC-10 allows for ratiometric analysis of mitochondrial membrane potential, where a shift from orange (Ex/Em: 540nm/590nm) to green fluorescence (Ex/Em: 490nm/525nm) is indicative of compromised mitochondria 5 x 50 μL
$248.00
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Packaging of mitochondrial activity assay kit, ION Vital MitoVolt
Mitochondrial membrane potential assay kit that uses JC-10, a dye that accumulates in polarized mitochondria where it emits a red-shifted fluorescence, that is more soluble than JC-1. 5 plates  
$308.00
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