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Evaluation of Intracellular Location of Reactive Oxygen Species in Solea Senegalensis Spermatozoa
Bu içeriği görüntülemek için JoVE aboneliği gereklidir. Oturum açın veya ücretsiz deneme sürümünü başlatın.
Evaluation of Intracellular Location of Reactive Oxygen Species in Solea Senegalensis Spermatozoa
Bölümler
- 00:00Başlık
- 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
- 07:49Sonuçlar
- 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.
