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Evaluering af intracellulær placering af reaktive ilt arter i Solea Senegalensis sædcellerne
JoVE Journal
Developmental Biology
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JoVE Journal Developmental Biology
Evaluation of Intracellular Location of Reactive Oxygen Species in Solea Senegalensis Spermatozoa
DOI:

10:24 min

March 11, 2018

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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

Summary

Automatic Translation

Denne protokol beskriver en detaljeret metode til påvisning af H2O2 lokalisering inden for Solea senegalensis sædcellerne ved hjælp af en følsom fluorokrom DCFH-DA for ROS, en levende mitokondrier pletten for mitokondrier og DAPI for kerner visualisering, henholdsvis. Protokollen er designet til at blive udført inden for 2 h med enten friske eller optøede sædcellerne.

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