The MTT assay and the MTS assay are laboratory tests and standard colorimetric assays (an assay which measures changes in color) for measuring the activity of enzymes that reduce MTT or MTS + PMS to formazan Formazan dyes are artificial chromogenic products of the reduction of tetrazolium salts by dehydrogenases and reductases. They have a variety of colors from dark blue to deep red to orange, depending on the original tetrazolium salt used as the substrate for the reaction, giving a purple color. It can also be used to determine cytotoxicity Cytotoxicity is the quality of being toxic to cells. Examples of toxic agents are a chemical substance, an immune cell or some types of venom of potential medicinal allMedicine is the art and science of healing. It encompasses a range of health care practices evolved to maintain and restore health by the prevention and treatment of illness agents and other toxic materials, since those agents would result in cell toxicity and therefore metabolic dysfunction and therefore decreased performance in the assay.

Yellow MTT (3-(4,5-Di Numerical prefixes are prefixes usually derived from the words for numbers in various languages, most commonly Greek and Latin, although this is not always the case methyl In chemistry, a methyl group is a hydrophobic alkyl functional group named after methane (C thiazol Thiazole, or 1,3-thiazole, is a clear to pale yellow flammable liquid with a pyridine-like odor and the molecular formula C3H3NS. It is a 5-membered ring, in which two of the vertices of the ring are nitrogen and sulfur, and the other three are carbons-2-yl)-2,5-diphenyl where the six carbon atoms are arranged in a cyclic ring structure. This hydrophobic, highly-stable and aromatic hydrocarbon unit can be found in many organic compounds. It can be thought of as being derived from benzene . In fact, in chemical literature benzene itself is sometimes denoted as PhHtetrazolium bromide, a tetrazole) is reduced to purple Purple is a general term used in English for the range of shades of color occurring between red and blue. In additive light combinations it occurs by mixing the primary colors red and blue in varying proportions. In subtractive pigments it can be equal to the primary color magenta or be formed by mixing magenta with the secondary colors red or formazan Formazan dyes are artificial chromogenic products of the reduction of tetrazolium salts by dehydrogenases and reductases. They have a variety of colors from dark blue to deep red to orange, depending on the original tetrazolium salt used as the substrate for the reaction in living cells.^[1] A solubilization solution (usually either dimethyl sulfoxide Dimethyl sulfoxide is the chemical compound with the formula (CH3)2SO. It was first synthesized in 1866 by the Russian scientist Alexander Zaytsev, who reported his findings in a German chemistry journal in 1867. This colorless liquid is an important polar aprotic solvent that dissolves both polar and nonpolar compounds and is miscible in a wide, an acidified ethanol solution, or a solution of the detergent A detergent is a material intended to assist cleaning. The term is sometimes used to differentiate between soap and other surfactants used for cleaning. As an adjective pertaining to a substance, it (or "detersive") means "cleaning" or "having cleaning properties"; "detergency" indicates presence or degree sodium dodecyl sulfate Sodium lauryl sulfate or sodium dodecyl sulfate (SDS or NaDS) (C12H25S in diluted hydrochloric acid Hydrochloric acid is the solution of hydrogen chloride (H ) is added to dissolve the insoluble purple formazan Formazan dyes are artificial chromogenic products of the reduction of tetrazolium salts by dehydrogenases and reductases. They have a variety of colors from dark blue to deep red to orange, depending on the original tetrazolium salt used as the substrate for the reaction product into a colored solution. The absorbance where I is the intensity of light at a specified wavelength λ that has passed through a sample and I0 is the intensity of the light before it enters the sample or incident light intensity. Absorbance measurements are often carried out in analytical chemistry, since the absorbance of a sample is proportional to the thickness of the sample and the of this colored solution can be quantified by measuring at a certain wavelength In physics, the wavelength of a sinusoidal wave is the spatial period of the wave – the distance over which the wave's shape repeats. It is usually determined by considering the distance between consecutive corresponding points of the same phase, such as crests, troughs, or zero crossings, and is a characteristic of both traveling waves and (usually between 500 and 600 nm) by a spectrophotometer In physics, spectrophotometry is the quantifiable study of electromagnetic spectra. It is more specific than the general term electromagnetic spectroscopy in that spectrophotometry deals with visible light, near-ultraviolet, and near-infrared. Also, the term does not cover time-resolved spectroscopic techniques. The absorption maximum is dependent on the solvent employed.

MTS is a more recent alternative to MTT. MTS (3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium), in the presence of phenazine methosulfate (PMS), produces a water-soluble formazan product that has an absorbance maximum at 490-500 nm in phosphate-buffered saline.^[2] It is advantageous over MTT in that (1) the reagents MTS + PMS are reduced more efficiently than MTT, and (2) the product is water soluble, decreasing toxicity to cells seen with an insoluble product.

These reductions Redox describes all chemical reactions in which atoms have their oxidation number (oxidation state) changed. This can be either a simple redox process such as the oxidation of carbon to yield carbon dioxide or the reduction of carbon by hydrogen to yield methane (CH4), or it can be a complex process such as the oxidation of sugar in the human body take place only when reductase enzymes Enzymes are proteins that catalyze chemical reactions. In enzymatic reactions, the molecules at the beginning of the process are called substrates, and the enzyme converts them into different molecules, called the products. Almost all processes in a biological cell need enzymes to occur at significant rates. Since enzymes are selective for their are active, and therefore conversion is often used as a measure of viable (living) cells. However, it is important to keep in mind that other viability tests (such as the CASY cell counting technology) sometimes give completely different results, as many different conditions can increase or decrease metabolic activity. Changes in metabolic activity can give large changes in MTT or MTS results while the number of viable cells is constant. When the amount of purple formazan produced by cells treated with an agent is compared with the amount of formazan produced by untreated control When an experiment is conducted for the purpose of determining the effect of a single variable of interest on a particular system, a scientific control is used to minimize the unintended influence of other variables on the same system. Such extraneous variables include researcher bias, environmental changes, and biological variation. Scientific cells, the effectiveness of the agent in causing death, or changing metabolism of cells, can be deduced through the production of a dose-response curve The dose-response relationship, or exposure-response relationship, describes the change in effect on an organism caused by differing levels of exposure to a stressor (usually a chemical) after a certain exposure time. This may apply to individuals (eg: a small amount has no observable effect, a large amount is fatal), or to populations (eg: how.

References

1. ^ Mosmann T (December 1983). "Rapid colorimetric assay for cellular growth and survival: application to proliferation and cytotoxicity assays". Journal of immunological methods 65 (1-2): 55–63. PMID A PMID is a unique number assigned to each PubMed citation of life sciences and biomedical scientific journal articles. The related Pubmed Central archive may additionally assign a separate number, a PMCID (PubMed Central Identifier), normally written with a PMC prefix 6606682. http://linkinghub.elsevier.com/retrieve/pii/0022-1759(83)90303-4.
2. ^ Cory AH, Owen TC, Barltrop JA, Cory JG (July 1991). "Use of an aqueous soluble tetrazolium/formazan assay for cell growth assays in culture". Cancer communications 3 (7): 207–12. PMID A PMID is a unique number assigned to each PubMed citation of life sciences and biomedical scientific journal articles. The related Pubmed Central archive may additionally assign a separate number, a PMCID (PubMed Central Identifier), normally written with a PMC prefix 1867954.

• Wilson, A. P., Cytotoxicity and Viability Assays in Animal Cell Culture: A Practical Approach, 3rd ed. (ed. Masters, J. R. W.) Oxford University Press: oXford 2000, Vol. 1,
• Mitochondrial and nonmitochondrial reduction of MTT: interaction of MTT with TMRE, JC-1, and NAO mitochondrial fluorescent probes. Bernas T, Dobrucki J. Cytometry. 2002 Apr 1;47(4):236-42. PMID 11933013

See also

• Tetrazolium chloride Triphenyl tetrazolium chloride, TTC, or simply Tetrazolium chloride is a redox indicator commonly used in biochemical experiments especially to indicate cellular respiration. It is a white crystalline powder, soluble in water, ethanol and acetone but insoluble in ether
• Formazan Formazan dyes are artificial chromogenic products of the reduction of tetrazolium salts by dehydrogenases and reductases. They have a variety of colors from dark blue to deep red to orange, depending on the original tetrazolium salt used as the substrate for the reaction MTT方法

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