title: Neutral red cytotoxicity assays for assessing in vivo carbon nanotube ecotoxicity in mussels--Comparing microscope and microplate methods.
abstract: The purpose of the present study was to compare two neutral red retention methods, the more established but very labour-intensive microscope method (NRR) against the more recently developed microplate method (NRU). The intention was to explore whether the sample volume throughput could be increased and potential operator bias avoided. Mussels Mytilus sp. were exposed in vivo to 50, 250 and 500 μg L(-1) single (SWCNTs) or multi-walled carbon nanotubes (MWCNTs). Using the NRR method, SWCNTs and MWCNTs caused concentration dependent decreases in neutral red retention time. However, a concentration dependent decrease in optical density was not observed using the NRU method. We conclude that the NRU method is not sensitive enough to assess carbon nanotube ecotoxicity in vivo in environmentally relevant media, and recommend using the NRR method.
authors.affiliation: Centre for Marine Biodiversity & Biotechnology, School of Life Sciences, Heriot-Watt University, Edinburgh, Scotland, UK.
authors.forename: M A
authors.initials: MA
authors.lastname: Miller
authors.affiliation: Centre for Marine Biodiversity & Biotechnology, School of Life Sciences, Heriot-Watt University, Edinburgh, Scotland, UK.
authors.forename: C
authors.initials: C
authors.lastname: Bankier
authors.affiliation: Centre for Marine Biodiversity & Biotechnology, School of Life Sciences, Heriot-Watt University, Edinburgh, Scotland, UK; King Abdulaziz University, Department of Biological Sciences, Faculty of Sciences, Jeddah, Saudi Arabia.
authors.forename: M A M
authors.initials: MA
authors.lastname: Al-Shaeri
authors.affiliation: Centre for Marine Biodiversity & Biotechnology, School of Life Sciences, Heriot-Watt University, Edinburgh, Scotland, UK. Electronic address: m.hartl@hw.ac.uk.
authors.forename: M G J
authors.initials: MG
authors.lastname: Hartl
mesh_terms: D000818:Animals; D001681:Biological Assay / Q000295:instrumentation / Q000379:methods; D049872:Bivalvia / Q000737:chemistry / Q000166:cytology / Q000187:drug effects*; D004305:Dose-Response Relationship, Drug; D054750:Ecotoxicology / Q000295:instrumentation / Q000379:methods*; D004784:Environmental Monitoring / Q000295:instrumentation / Q000379:methods*; D006458:Hemolymph / Q000737:chemistry / Q000166:cytology / Q000187:drug effects; D008247:Lysosomes / Q000737:chemistry / Q000187:drug effects; D046529:Microscopy, Electron, Transmission; D008962:Models, Theoretical; D037742:Nanotubes, Carbon / Q000737:chemistry / Q000633:toxicity*; D009499:Neutral Red / Q000032:analysis*; D012623:Seawater / Q000737:chemistry; D012680:Sensitivity and Specificity; D013053:Spectrophotometry
chemical_list: D037742:Nanotubes, Carbon; D009499:Neutral Red
keywords: Carbon nanotubes; Cytotoxicity; Ecotoxicity; Mytilus; Neutral red