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| The colour change of the indicator is a sensitive marker for the mitochondrial activity in the cell. |
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The mitochondrial activity in the cell, is considered as a measure of the zytotoxical effect. The picture shows tissues in the 24 well plate photographed from above, after the MTT has been metabolised. Violet tissue areas were metabolically active and have transformed MTT into its coloured formazan salt. In the next step the insoluble formazan salt is extracted with isopropanol and the absorbtion of the isopropanol solution is quantified at 570nm.
Succinate dehydrogenase : Activity measurement (MTT-Assay)
Principle:
The enzyme is localised in the mitochondria, and changes the yellow, water soluble dye diphenyltetrazoliumbromide (MTT) into a violet, insoluble product (MTT-formazan).
Implementation:
At the end of the incubation with the test substance, the samples (cells, tissue) are incubated in medium with MTT during 10 – 60 minutes at 37 degrees celsius. The dye enriched in the cells is released with an alcoholic solution, which is measured at 570nm in a microplate reader. The amount of released dye is related to the mitochondrial activity and thus the metabolic activity in the cells. This can (usually), but must not, correlate with the number of living cells. For this reason the results must be verified with those of the MTT test (i.e those with a second end point).
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