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Immunostaining of Whole-Mount Drosophila Testes for 3D Confocal Analysis of Large Spermatocytes
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Biologia
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JoVE Journal Biologia
Immunostaining of Whole-Mount Drosophila Testes for 3D Confocal Analysis of Large Spermatocytes

Immunostaining of Whole-Mount Drosophila Testes for 3D Confocal Analysis of Large Spermatocytes

DOI:
10.3791/61061-v

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08:37 min

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August 27, 2020

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, ,
1CNRS, Institut Jacques Monod,Université de Paris, F 75006, Paris, France

Capítulos

  • 00:05Introduction
  • 00:47Drosophila Testes Collection
  • 02:14Paraformaldehyde Fixation
  • 03:11Antibody Staining
  • 06:11Results: Representative Whole-Mount Drosophila Testis Immunostaining and 3D Confocal Analysis
  • 07:54Conclusion

Summary

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We present here an improved protocol for the whole-mount preparation and immunostaining of Drosophila testes, suitable for confocal microscopy and also allowing reproducible and reliable labeling. We illustrate this protocol by 3D representation and quantification of colocalization experiments on the large S4-S5 spermatocytes.

Tags

Whole-mount ImmunostainingDrosophila Testes3D Confocal AnalysisSpermatocytesNP40HeptaneFixation ProtocolAntibody PenetrationQuantifiable LabelingDiploid Germ Cell EvaluationRinger’s BufferParaformaldehydePBS-TPermeabilizationBlocking3D Imaging

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