Cell viability, cell proliferation, and cytotoxicity assays
Cell viability assays measure various cell markers, such as ATP or metabolic enzyme activity, as indicators of cell health. Many assays for cell viability and cell proliferation, including a variety of cytotoxicity assays, are detected using microplate readers. Viability assays using different detection modes are available, each with its own advantages.
What is cell viability?
Cell viability refers to the number of healthy cells in a population and can be evaluated using assays that measure enzyme activity, cell membrane integrity, ATP production, and other indicators. These methods can employ luminescent, fluorescent, or colorimetric readouts as indicators of general cell viability or even specific cellular pathways. Cytotoxicity and cell viability assays are often used to assess a drug or other treatment’s effect and are valuable tools in the search for new therapeutics, as well as advancing our understanding of how normal cells function.
What is cell proliferation?
Cell proliferation refers to the overall increase in cell number and is vital for tissue growth. It is the result of both cell growth and cell division. Both processes are highly regulated in healthy organisms to balance cellular size and population size. In experimental setups, monitoring cell proliferation can give insights into the effects of drugs or other treatments on a population of cells.
Featured cellular health resources
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Access cytotoxicity using the Delfia cell proliferation assay
Cell proliferation is an important parameter to measure when testing compound toxicity or the effect of cytokines on a population of cells. Since cell proliferation requires the synthesis of DNA, one accurate way to measure cell proliferation is to monitor the uptake of the modified thymidine nucleotide BrdU1. The DELFIA® cell proliferation assay is a time-resolved immunoassay based on the incorporation of BrdU into newly synthesized DNA strands of proliferating cells.
In this application note, we used the DELFIA cell proliferation assay to assess cytotoxicity of the chemotherapy drugs paclitaxel and etoposide in HeLa cells using the SpectraMax® i3 Multi-Mode Microplate Reader.
Application note: Delfia cell proliferation assay on the SpectraMax i3 Multi-Mode Microplate Reader
ATP-based cell viability assays
Luminescent ATP assays for cell viability and cytotoxicity provide a sensitive and rapid way to measure the number of viable cells in culture and quantify the cytotoxic effects of experimental treatments. These assays can be done in a high-throughput format using SpectraMax® microplate readers.
Learn about the CellTiter-Glo Luminescent Cell Viability Assay and more:
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Colorimetric cell viability, cell proliferation, cytotoxicity assays (MTT, XTT, MTS)
Colorimetric assays using tetrazolium salts like MTT, XTT, and MTS can be used to measure cell proliferation and cytotoxicity. Enzymes present in viable cells reduce a chromogenic reagent to form a colored product that can be quantified by measuring the absorbance.
Read the application note to learn about some of these methods:
EarlyTox Cell Viability Assay Kits
Cell viability assays are critical to a broad spectrum of research areas ranging from investigation into the mechanisms of cell death to the development of new therapeutics targeting apoptosis in diseases. One of the most popular detection technologies for cell viability is a fluorescence microplate reader.
Here are a few applications you may find of interest:
- Measuring cell health on the SpectraMax iD3 reader with cell viability assays
- EarlyTox Live/Dead Assay Kit on SpectraMax Fluorescence Microplate Readers
- EarlyTox Caspase-3/7 R110 Assay Kit on SpectraMax Fluorescence Microplate Readers
- EarlyTox Glutathione Assay Kit on SpectraMax Fluorescence Microplate Readers
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Measure oxidative metabolism and glycolytic activity
To survive, cells require fuel in the form of ATP to carry out most essential bioprocesses. This fuel is generated through glycolysis and mitochondrial respiration. While both produce ATP, glycolysis can function in the absence of oxygen, whereas mitochondria require oxygen for the final step in oxidative phosphorylation (OXPHOS). Understanding how these pathways respond to effector compounds can provide useful insights into the overall function of cells and the underlying mechanisms that determine cell fate.
Read our application note to learn about real-time data acquisition and analysis of the progression of oxygen consumption and glycolic activity:
Measuring cell proliferation using the CyQUANT Kit
Quantitation of cell proliferation using fluorescence allows one to easily monitor the effects of drugs and other experimental treatments on cell growth. The CyQUANT Cell Proliferation Assay Kit from Life Technologies is a sensitive, rapid and convenient way to quantitate cell growth using a fluorescence microplate reader.
This application note describes how to use the CyQUANT kit with SpectraMax® Microplate Readers and SoftMax® Pro Software from Molecular Devices.
Application note: Measuring cell proliferation using the CyQUANT Cell Proliferation Assay with SpectraMax Microplate Readers
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Reactive Oxygen Species (ROS)
Reactive Oxygen Species (ROS) are chemically reactive molecules containing oxygen. In eukaryotes, these molecules are mainly created during aerobic respiration and can cause problems, such as DNA damage and lipid peroxidation, resulting in cellular damage.
We demonstrate how SpectraMax microplate readers can be used to accurately quantify ROS levels using a luminescence-based assay:
Toxicology
Toxicology is the study of adverse effects of natural or man-made chemicals on living organism. It is a growing concern in our world today as we are exposed to more and more chemicals, both in our environment and in the products we use.
Latest Resources
Resources of Cell health: Viability, proliferation, cytotoxicity, and cellular function
Application Note
Understanding organoid morphology: a study to assess organoid size and cell count from 3D image analysis
Understanding organoid morphology: a study to assess organoid size and cell count from 3D image analysis
Compound library screens using 2D monolayer cancercell lines are commonly performed in the pharmaceutical industry, to identify molecules with therapeutic potential. Multiple analogues and…
Application Note
Spectral Fusion™ Illumination technology for an extended dynamic range on the SpectraMax i3x Multi-Mode Microplate Reader
Spectral Fusion™ Illumination technology for an extended dynamic range on the SpectraMax i3x Multi-Mode Microplate Reader
Many microplate readers and spectrophotometers use a photomultiplier tube (PMT) to detect fluorescence and luminescence signals. PMTs work by initially detecting photons from the sample…
Scientific Poster
Automation for organoid assays – An integrated system with high-content imaging
Automation for organoid assays – An integrated system with high-content imaging
Three-dimensional (3D) cell models that represent various tissues are being successfully used in drug discovery and disease modeling to study complex biological effects and tissue architectu…
Application Note
Rapidly assess drug response in 2D and 3D breast cancer model systems with a luminescent viability assay
Rapidly assess drug response in 2D and 3D breast cancer model systems with a luminescent viability assay
For many years, two-dimensional (2D) cell culture involving cells grown in a monolayer on a flat culture surface has served as a convenient system for investigating disease mechanisms and…
Application Note
CloneSelect Imager FL fluorescence for rapid day zero monoclonality
CloneSelect Imager FL fluorescence for rapid day zero monoclonality
Gene editing allows DNA manipulation, i.e., addition, deletion, or modification using different tools. Several nucleases for genome editing are now available: zinc finger nucleases (ZFN),…
Application Note
Detect endotoxin with the Kinetic-QCL Kinetic Chromogenic LAL Assay
Detect endotoxin with the Kinetic-QCL Kinetic Chromogenic LAL Assay
Monitoring for contaminants is a critical step during the production process in the pharmaceutical and medical device industries. A frequent contaminant, endotoxin, can cause fever,…
Blog
How automated organoid cell cultures are developed, imaged, and analyzed
How automated organoid cell cultures are developed, imaged, and analyzed
3D cell models are becoming increasingly popular for studying complex biological effects, tissue functionality, and diseases. Their ability to self-organize and mimic…
eBook
The Ultimate Guide to Microplate Reader Solutions
The Ultimate Guide to Microplate Reader Solutions
Evaluating microplate readers doesn’t have to be overwhelming. First, consider your application needs. If your budget is modest, a single-mode reader dedicated to your main application may b…
Publications
Multiple Steps from Microplate-Reader Workstations
Multiple Steps from Microplate-Reader Workstations
Scientists take many roads to overcoming bottlenecks in workflows. For example, a workstation including a microplate reader can handle multiple steps, which improves efficiency and adds spee…
Application Note
Measure cancer cell viability using a homogeneous, stable luminescence assay
Measure cancer cell viability using a homogeneous, stable luminescence assay
Luminescent cell viability assays offer sensitivity and an easy workflow for monitoring the effects of various experimental conditions.
Application Note
Assess cell viability and proliferation with colorimetric readouts
Assess cell viability and proliferation with colorimetric readouts
Learn how absorbance microplate reader provides a colorimetric readouts for cell viability or proliferation.
Application Note
Measure oxidative metabolism and glycolytic activity on the SpectraMax i3x reader
Measure oxidative metabolism and glycolytic activity on the SpectraMax i3x reader
Learn how to measure oxidative metabolism and glycolytic activity on the SpectraMax i3x reader using assays from Agilent Technologies.
Application Note
Measuring cell health on the SpectraMax iD3 reader with cell viability assays
Measuring cell health on the SpectraMax iD3 reader with cell viability assays
Measuring cell health from the multiple vantage points of viability and apoptotic pathways can provide insight into the impacts of a variety of experimental treatments including drug…
Application Note
Measuring reactive oxygen species with SpectraMax microplate readers
Measuring reactive oxygen species with SpectraMax microplate readers
Reactive Oxygen Species (ROS) are chemically reactive molecules containing oxygen. In eukaryotes, these molecules are mainly created during aerobic respiration and can cause problems, such…
Application Note
EarlyTox Glutathione Assay Kit on SpectraMax Fluorescence Microplate Readers
EarlyTox Glutathione Assay Kit on SpectraMax Fluorescence Microplate Readers
Apoptosis is a highly regulated cellular program that causes cell death in normal processes such as embryonic development, as well as diseases including cancer and neurodegenerative…
Application Note
EarlyTox Live/Dead Assay Kit on SpectraMax Fluorescence Microplate Readers
EarlyTox Live/Dead Assay Kit on SpectraMax Fluorescence Microplate Readers
Cell viability assays are often performed to assess the effects of a variety of treatments, including drug candidates, pathway activators and inhibitors, and reporter genes. One of the most…
Application Note
Luminescent cell viability and cytotoxicity assays on the SpectraMax i3x Multi-Mode Microplate Reader
Luminescent cell viability and cytotoxicity assays on the SpectraMax i3x Multi-Mode Microplate Reader
The CellTiter-Glo assay from Promega uses luciferase enzyme, which requires ATP in order to generate light. The luminescent signal produced in the assay depends on the amount of ATP in the…
Application Note
EarlyTox Caspase-3/7 R110 Assay Kit on SpectraMax Fluorescence Microplate Readers
EarlyTox Caspase-3/7 R110 Assay Kit on SpectraMax Fluorescence Microplate Readers
Apoptosis is a highly regulated cellular program that causes cell death in normal processes such as embryonic development, as well as diseases including cancer and neurodegenerative…