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Mtt lecture
Mtt lecture
Mtt lecture
Mtt lecture
Mtt lecture
Mtt lecture
Mtt lecture
Mtt lecture
Mtt lecture
Mtt lecture
Mtt lecture
Mtt lecture
Mtt lecture
Mtt lecture
Mtt lecture
Mtt lecture
Mtt lecture
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Mtt lecture

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  • Viable cells that active mitochondria reductase which produce the purple formazan crystals\n
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  • Transcript

    • 1. MTT Assay
    • 2. MTT AssayPurpose: To understand how theMTT asay works, what it measures, and why it is important.
    • 3. How it works:The MTT assay is a colorimetric assay systemand is a quantitative method used to determinecell growth and viability.
    • 4. Why is the MTT assay important: measuring cellular proliferation (cell growth) in response to external factors
    • 5. Applications:•Drug sensitivity•Cytotoxicity•Response to growth factors•Cell activation
    • 6. Yellow MTT (3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide, atetrazole) is reduced to purpleformazan in the mitochondria ofliving cells. (Formazan dyes areartificial chromogenic substrates fordehydrogenases and reductases).
    • 7. In mitochondria :
    • 8. - + - - + + ++ + + ++ + ++ + + + - + - + + - + - + + + + + + ++ + + + + + + - + + + + + + + + -+ - + + + + + + + + + + + + + + - + -+ + + + - - + ++ + ++ + + ++ - + + + +
    • 9. Viable Cell -+ + + + + + ++ - + + + + + -+ - + + + - + + + + + + + +
    • 10. Non-viable Cell - + - + + - + - + + + + + + ++ + + + + + + - + + + + + + + + -+ - + + + + + + + + + + + + + + - + -+ + + + - - + ++ + ++ + + ++ - + + + +
    • 11. DMSO+-++ ++ +
    • 12. Actinomycin D is a substance that binds to DNAand prevents transcription.In this assay it serves as a negative control byinhibiting WEHI cell replication during theincubation time and ensures maximum assaysensitivity.
    • 13. Count Adjust count cells to 2x105cells/ mL WEHI cells, Mɸ cells, use use RPMI- RPMI-1640 WEHI WEHI+ActD
    • 14. Wednesday Before Lecture MTT10 15 10µµl µl l
    • 15. DMSOToday 100 µl
    • 16. Today Read Incubate for absorbance 5’ in the at 570nm dark

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