Resazurin sodium salt

Assessment of Cell Viability and Cytotoxicity Using Resazurin Sodium Salt^[1]

This experimental protocol is compiled based on research content and is for reference only; please adjust according to actual needs.

(1) Reagent preparation: Prepare a stock solution (400μM) of Resazurin sodium salt in double-distilled water.

(2) Cell seeding: Seed 2–8×10³ viable cells per well in a 96-well standard tissue culture plate (200μL/well); incubate overnight at 37°C, 5% CO₂, saturated humidity.

(3) Sample pretreatment: Tap water samples must first be vacuum-filtered through a 0.15μm polytetrafluoroethylene (PTFE) microporous membrane to remove particulate matter.

(4) Resazurin sodium salt assay: Add 20μL of Resazurin sodium salt solution (final concentration 40μM) to each well, return to 37°C, 5% CO₂ for continued incubation, and read fluorescence (Ex=530-560nm, Em=590nm) with a microplate reader at predetermined time points after addition.

Precautions:

(1) Resazurin sodium salt may chemically react with some test compounds; perform a cross-reaction verification in a cell-free system before formal experiments.

(2) The reduction rate of Resazurin sodium salt varies significantly among different cell lines; optimize the Resazurin sodium salt concentration and incubation time to avoid underestimating cell viability.

(3) For your safety and health, wear laboratory coat and disposable gloves during operation.

References:
[1] O'Brien J, Wilson I, Orton T, et al. Investigation of the Alamar Blue (resazurin) fluorescent dye for the assessment of mammalian cell cytotoxicity. Eur J Biochem. 2000 Sep;267(17):5421-6.