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Mtt lecture

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The MTT assay is a quantitative method to measure cell growth and viability. It works by using the yellow tetrazolium dye MTT, which is reduced to purple formazan in the mitochondria of living cells. The amount of formazan produced is proportional to the number of viable cells and can be measured spectrophotometrically. The MTT assay is important for measuring cellular proliferation in response to external factors and has applications in testing drug sensitivity, cytotoxicity, and cell responses to growth factors.

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MTT Assay
MTT Assay Purpose: To understand how the MTT asay works, what it measures, and why it is important.
How it works: The MTT assay is a colorimetric assay system and is a quantitative method used to determine cell growth and viability.
Why is the MTT assay important: measuring cellular proliferation (cell growth) in response to external factors
Applications: Drug sensitivity Cytotoxicity Response to growth factors Cell activation
Yellow MTT (3-(4,5- Dimethylthiazol-2-yl)-2,5- diphenyltetrazolium bromide, a tetrazole) is reduced to purple formazan in the mitochondria of living cells. (Formazan dyes are arti鍖cial chromogenic substrates for dehydrogenases and reductases).
In mitochondria :
- + - - + + + + + + + + + + + + + + - + - + + - + - + + + + + + + + + + + + + + - + + + + + + + + -+ - + + + + + + + + + + + + + + - + -+ + + + - - + + + + + + + + + + - + + + +
Viable Cell -+ + + + + + + + - + + + + + - + - + + + - + + + + + + + +
Non-viable Cell - + - + + - + - + + + + + + + + + + + + + + - + + + + + + + + -+ - + + + + + + + + + + + + + + - + -+ + + + - - + + + + + + + + + + - + + + +
DMSO +-+ + + + +
Actinomycin D is a substance that binds to DNA and prevents transcription. In this assay it serves as a negative control by inhibiting WEHI cell replication during the incubation time and ensures maximum assay sensitivity.
Count Adjust count cells to 2x105cells/ mL WEHI cells, M標 cells, use use RPMI- RPMI-1640 WEHI WEHI+ActD
Wednesday Before Lecture MTT 10 15 10袖 袖l 袖l l
DMSO Today 100 袖l
Today Read Incubate for absorbance 5 in the at 570nm dark
Mtt lecture

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Mtt lecture

  • 2. MTT Assay Purpose: To understand how the MTT asay works, what it measures, and why it is important.
  • 3. How it works: The MTT assay is a colorimetric assay system and is a quantitative method used to determine cell growth and viability.
  • 4. Why is the MTT assay important: measuring cellular proliferation (cell growth) in response to external factors
  • 6. Yellow MTT (3-(4,5- Dimethylthiazol-2-yl)-2,5- diphenyltetrazolium bromide, a tetrazole) is reduced to purple formazan in the mitochondria of living cells. (Formazan dyes are arti鍖cial chromogenic substrates for dehydrogenases and reductases).
  • 8. - + - - + + + + + + + + + + + + + + - + - + + - + - + + + + + + + + + + + + + + - + + + + + + + + -+ - + + + + + + + + + + + + + + - + -+ + + + - - + + + + + + + + + + - + + + +
  • 9. Viable Cell -+ + + + + + + + - + + + + + - + - + + + - + + + + + + + +
  • 10. Non-viable Cell - + - + + - + - + + + + + + + + + + + + + + - + + + + + + + + -+ - + + + + + + + + + + + + + + - + -+ + + + - - + + + + + + + + + + - + + + +
  • 12. Actinomycin D is a substance that binds to DNA and prevents transcription. In this assay it serves as a negative control by inhibiting WEHI cell replication during the incubation time and ensures maximum assay sensitivity.
  • 13. Count Adjust count cells to 2x105cells/ mL WEHI cells, M標 cells, use use RPMI- RPMI-1640 WEHI WEHI+ActD
  • 14. Wednesday Before Lecture MTT 10 15 10袖 袖l 袖l l
  • 15. DMSO Today 100 袖l
  • 16. Today Read Incubate for absorbance 5 in the at 570nm dark

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