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Evaluering av intracellulær plasseringen av reaktive oksygen arter i Solea Senegalensis Spermatozoa
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 protokollen beskriver en detaljert metodikk for å oppdage H2O2 lokalisering i Solea senegalensis spermatozoa bruker en følsom fluorochrome DCFH-DA for ROS, en live mitokondrier flekk for mitokondrier og DAPI for kjerner visualisering, henholdsvis. Protokollen er utformet som skal utføres innen 2 timer med enten friske eller tinte spermatozoa.

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