Nanotechnologies. 5-(and 6)-Chloromethyl-2’,7’ Dichlorodihydrofluorescein diacetate (CM-H2DCF- DA) assay for evaluating nanoparticle-induced intracellular reactive oxygen species (ROS) production in RAW 264.7 macrophage cell line

Nanotechnologies. 5-(and 6)-Chloromethyl-2’,7’ Dichlorodihydrofluorescein diacetate (CM-H2DCF- DA) assay for evaluating nanoparticle-induced intracellular reactive oxygen species (ROS) production in RAW 264.7 macrophage cell line

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1   Scope

This document describes how to test and evaluate results obtained from in vitro ROS generation in RAW 264.7 macrophage cells exposed to nano-objects, nanoparticles, their aggregates and agglomerates using the CM-H2DCFDA assay.
The protocol in this document is limited to use of a 24 well plate so if other plates were to be used, volumes would need to be adjusted and the protocol steps validated to ensure confidence in the test results.