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 shop-only through careful design

through careful design

All SAE shall comply with the safety requirements and/or protective measures in Clause 6 to Clause 14 of BS EN 1808

the measurements can be used for comparison purposes

The registered object identifiers to which this part of ISO/IEC 13522 refers are those used in ISO/IEC 13522-1

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 shop-only through careful design1 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.