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MTT Assay

A colorimetric assay for assessing cell metabolic activity. It is primarily used to measure cell proliferation, cell viability, and cytotoxicity. This assay is based on the reduction of the MTT reagent, a yellow tetrazolium salt, to formazan, a purple-colored insoluble product, by mitochondrial enzymes in living cells. The amount of Formazan produced is directly proportional to the number of viable cells.

Workflows requiring a MTT Assay

Cell Proliferation Studies

The MTT assay is integral to cell proliferation studies, a key area in basic biological research and cancer research. It provides quantitative insights into the growth rate of cells under diverse conditions, such as genetic modifications or exposure to growth factors, which is vital in understanding cancer cell dynamics and developing therapeutic strategies. In tissue engineering and regenerative medicine, this assay is essential for monitoring cell viability and growth within artificial scaffolds, thereby informing the design and efficacy of regenerative treatments.

Drug Efficacy and Toxicity Testing

In the realm of pharmaceutical research, the MTT assay is a cornerstone for evaluating new drug compounds, serving a dual role in assessing both efficacy and toxicity. It enables researchers to determine the therapeutic index by exposing cultured cells to various drug concentrations, thereby identifying optimal dosages that maximize efficacy while minimizing cytotoxic effects. This assay is particularly crucial in early drug development stages, guiding the selection of promising compounds and shaping the understanding of safe dosage limits and potential side effects.

Assessment of Cellular Response to Environmental Stress

In environmental toxicology, the MTT assay is employed to gauge cellular health in response to various stressors like pollutants or radiation. This application is crucial for evaluating the risks posed by environmental contaminants and understanding cellular defense mechanisms. Additionally, the assay is used in studies of cellular metabolism and physiology to explore how cells adapt to changes in their environment, such as nutritional variations or hypoxia, providing valuable insights into cellular resilience and survival strategies.


Methods of Performing a MTT Assay

Standard MTT Assay Protocol

The Standard MTT Assay Protocol is the most commonly used method, involving the direct application of MTT to cells and subsequent measurement of cell viability based on formazan production. This method is straightforward and is widely used for its simplicity and effectiveness in a variety of cell types.



High-Throughput MTT Assay

The High-Throughput MTT Assay is a modified version of the standard protocol, adapted for large-scale screening, such as in drug discovery. This method utilizes automated systems and is designed to handle a large number of samples efficiently.


MTT Assays have never been easier

The OT-2 is a bench-top liquid handler designed to be accessible and flexible enough to automate many common applications.

MTT Assay Combined with Complementary Techniques

This method involves combining the MTT assay with other analytical techniques, such as flow cytometry or microscopy, to provide additional insights into cell viability and morphology.


How to Automate the MTT Assay process:


Benefits of Automation over Manual Pipetting for MTT Assay:

Protocol Spotlights

Opentrons helps you automate MTT Assay with open-source protocols for the OT-2 and Opentrons Flex


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