MTT Test and Time-lapse Microscopy to Evaluate the Antitumor Potential of Nucleoside Analogues

Anticancer Res. 2021 Jan;41(1):137-149. doi: 10.21873/anticanres.14759.


Background/aim: Conventional viability tests, help to screen the cellular effects of candidate molecules, but the endpoint of these measurements lacks sufficient information regarding the molecular aspects. A non-invasive, easy-to-setup live-cell microscopic method served to in-depth analysis of mechanisms of potential anticancer drugs.

Materials and methods: The proposed method combining the MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) test with time-lapse scanning microscopy (TLS), provided additional data related to the cell-cycle and the dynamic properties of cell morphology. Apoptotic and necrotic events became detectable with these methods.

Results: Quantification of the results was assisted by image analysis of the acquired image sequences. After demonstrating the potential of the TLS method, a series of experiments compared the in vitro effect of a known and a newly synthesized nucleoside analogue.

Conclusion: The proposed approach provided a more in-depth insight into the cellular processes that can be affected by known chemotherapeutic agents including nucleoside analogues rather than applying repeated individual treatments.

Keywords: Antimetabolites; cellular morphology; nucleoside analogues; time-lapse microscopy.

MeSH terms

  • Antineoplastic Agents / pharmacology*
  • Cell Cycle / drug effects
  • Cell Death / drug effects
  • Cell Division / drug effects
  • Cell Line, Tumor
  • Cell Survival / drug effects
  • Cells, Cultured
  • Dose-Response Relationship, Drug
  • Humans
  • Microscopy
  • Nucleosides / analogs & derivatives
  • Nucleosides / pharmacology*
  • Tetrazolium Salts*
  • Thiazoles*
  • Time-Lapse Imaging* / methods


  • Antineoplastic Agents
  • Nucleosides
  • Tetrazolium Salts
  • Thiazoles
  • thiazolyl blue