/
/
Evaluation of Intracellular Location of Reactive Oxygen Species in Solea Senegalensis Spermatozoa
A subscription to JoVE is required to view this content. Sign in or start your free trial.
Chapters
- 00:00Title
- 01:32Preparatory work before the experiment
- 02:58Sample preparation before fluorochromes incubation
- 04:40Fluorochromes incubation
- 05:33Sample preparation for confocal microscopy
- 05:51Confocal setting up prior to the experiment
- 06:49Acquisition of images
- 07:49Results
- 08:24Creating a 3D image video
- 09:53Conclusions
This protocol describes a detailed methodology for detecting H2O2 localization within Solea senegalensis spermatozoa using a sensitive fluorochrome DCFH-DA for ROS, a live mitochondria stain for mitochondria, and DAPI for nuclei visualization, respectively. The protocol is designed to be performed within 2 h with either fresh or thawed spermatozoa.
