Resazurin Cytotoxicity Assay


Resazurin (7-Hydroxy-3H-phenoxazin-3-one 10-oxide), is a mildly fluorescent blue that that, upon reduction, becomes the highly fluorescent pink resorufin. The irreversible reduction of resazurin to resorcin is proportional to aerobic respiration. This is useful for detecting the viability of bacteria and mammalian cells by their redox capabilities.

Procedure (for 384-well plate)

  • Make 500 µM resazurin stock solution in PBS (stored at -20 or 4C).

-If storing at -20, reduce number of freeze/thaws by 10.

  • Filter-sterilize solution through a 0.2 µm filter into a sterile, light protected container.
  • Create a working solution of 10 µM from the resazurin stock solution
  • Prepare 384-well plates with cells and compound to have final well volume of around 25 µL/well. (See tissue-culture)
  • Incubate cells at 37C for desired amount of time.
  • Add 10 µL/well of 10 µM resazurin solution to cells.
  • Incubate plates 1-4 hours at 37C
  • Read fluorescence at 560 nm excitation and 590 nm emission.
Unless otherwise stated, the content of this page is licensed under Creative Commons Attribution-ShareAlike 3.0 License