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Quantitative Analysis of Protein Expression to Study Lineage Specification in Mouse Preimplantation Embryos
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发育生物学
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JoVE 杂志 发育生物学
Quantitative Analysis of Protein Expression to Study Lineage Specification in Mouse Preimplantation Embryos
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Quantitative Analysis of Protein Expression to Study Lineage Specification in Mouse Preimplantation Embryos

DOI:
10.3791/53654-v

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11:25 min

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February 22, 2016

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, , , ,
1Developmental Biology Program,Sloan Kettering Institute

Chapters

  • 00:05Title
  • 00:41Embryo Collection
  • 03:25Confocal Imaging
  • 04:47Image Analysis and Data Pre-processing
  • 08:44Results: Lineage Specification Analysis in Mouse Preimplantation Embryos
  • 10:30Conclusion

Summary

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This protocol presents a method to perform quantitative, single-cell in situ analysis of protein expression to study lineage specification in mouse preimplantation embryos. The procedures necessary for collection of blastocysts, whole-mount immunofluorescent detection of proteins, imaging of samples on a confocal microscope, and nuclear segmentation and image analysis are described.

Tags

Quantitative AnalysisProtein ExpressionLineage SpecificationMouse Preimplantation EmbryosConfocal MicroscopySingle-cell ResolutionSemi-automatedNuclear DensityCell Population HeterogeneityZona Pellucida RemovalImmunofluorescenceConfocal Imaging

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