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Use of Single Molecule Fluorescent In Situ Hybridization (SM-FISH) to Quantify and Localize mRNAs in Murine Oocytes
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Journal JoVE Génétique
Use of Single Molecule Fluorescent In Situ Hybridization (SM-FISH) to Quantify and Localize mRNAs in Murine Oocytes

Use of Single Molecule Fluorescent In Situ Hybridization (SM-FISH) to Quantify and Localize mRNAs in Murine Oocytes

DOI:
10.3791/59414-v

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08:18 min

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April 24, 2019

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,
1Department of Animal Science,University of Nebraska-Lincoln

Chapitres

  • 00:04Titre
  • 00:38Collection of Ovulated Oocytes from Female Mice
  • 02:06SM-FISH Staining of Oocytes
  • 04:21Image Processing
  • 06:21Results: mRNA Quantification Results using the Spot Finding and Tracking Program
  • 07:45Conclusion

Summary

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Traduction automatique

To reproducibly count the numbers of mRNAs in individual oocytes, single molecule RNA fluorescence in situ hybridization (RNA-FISH) was optimized for non-adherent cells. Oocytes were collected, hybridized with the transcript specific probes, and quantified using an image quantification software.

Tags

Keywords: Single Molecule Fluorescent In Situ Hybridization (SM-FISH)MRNA QuantificationMRNA LocalizationMurine OocytesNanogPou5f1DapBOocyte PreparationFixationPermeabilizationProtease TreatmentProbe Hybridization

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