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

Available format(s)

Hardcopy , PDF

Language(s)

English

Published date

31-08-2016

Publisher

British Standards Institution

Foreword
Introduction
1 Scope
2 Normative references
3 Terms and definitions
4 Symbols and abbreviated terms
5 Materials
6 Technical equipment
7 Nanoparticle sample preparation
8 Preparations
9 Evaluation of nanoparticle impact on ROS generation in cells
10 Data analysis and results
Annex A (informative) - Alternate cell lines
Annex B (informative) - Alternate fluorescence characterization
techniques
Annex C (informative) - Suspension preparation and characterization
Annex D (informative) - Example experimental data from RAW 264.7
Bibliography

Defines 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-H[2]DCFDA assay.

Committee	NTI/1
DocumentType	Standard
Pages	32
PublisherName	British Standards Institution
Status	Current

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.

Standards	Relationship
ISO/TS 19006:2016	Identical

ISO 29701:2010	Nanotechnologies Endotoxin test on nanomaterial samples for in vitro systems Limulus amebocyte lysate (LAL) test
ISO/TS 18827:2017	Nanotechnologies — Electron spin resonance (ESR) as a method for measuring reactive oxygen species (ROS) generated by metal oxide nanomaterials
ISO/TR 13097:2013	Guidelines for the characterization of dispersion stability
ISO/TS 80004-2:2015	Nanotechnologies — Vocabulary — Part 2: Nano-objects

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PD ISO/TS 19006:2016

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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Abstract

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