Neobenedenia sp.

Trujillo-Gonzalez ́, Alejandro, Constantinoiu, Constantin C., Rowe, Richard & Hutson, Kate S., 2015, Tracking transparent monogenean parasites on fish from infection to maturity, International Journal for Parasitology: Parasites and Wildlife 4 (3), pp. 316-322 : 317

2.2. Fluorescent labelling of Neobenedenia sp. oncomiracidia

Neobenedenia sp. oncomiracidia were labelled with a fluorescent marker to identify individual parasites on the fish body surface. A 10 mM stock solution of the fluorescent dye 5(6)-carboxyfluorescein diacetate N-succinimidyl ester (CFSE; Sigma ‾ Aldrich, Castle Hill, NSW, Australia) was made by resuspending CFSE lyophilised powder in 100% dimethyl sulphoxide (DMSO), and stored at 4 ǫ C in dark conditions until use. The stock solution was diluted with filtered sea water (35 ppt) to produce a 30 nM working solution of CFSE for labelling (modified from Glennon et al., 2007). Approximately 400 Neobenedenia sp. oncomiracidia were held for 15 min in dark conditions in a 50 mL beaker with 25 mL of sea water (35 ppt) and 5 mL of 30 nM CFSE working solution. Only swimming oncomiracidia were selected for the experiments.