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Imaging- and Flow Cytometry-based Analysis of Cell Position and the Cell Cycle in 3D Melanoma Spheroids
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JoVE Journal Medicine
Imaging- and Flow Cytometry-based Analysis of Cell Position and the Cell Cycle in 3D Melanoma Spheroids

Imaging- and Flow Cytometry-based Analysis of Cell Position and the Cell Cycle in 3D Melanoma Spheroids

DOI:
10.3791/53486-v

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10:44 min

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December 28, 2015

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, , , ,
1The Centenary Institute, 2Sydney Medical School,University of Sydney, 3The University of Queensland Diamantina Institute, Translational Research Institute,The University of Queensland, 4Department of Dermatology,Royal Prince Alfred Hospital, 5Discipline of Dermatology,University of Sydney

Chapters

  • 00:05Title
  • 01:063D Spheroid Formation
  • 03:30Image Analysis of FUCCI Spheroid Sections
  • 08:40Results: Quantification of G1 Arrested and Proliferating Cells in Cancer Cell Spheroids
  • 10:00Conclusion

Summary

Automatic Translation

Automatic Translation

We describe two complementary methods using the fluorescence ubiquitination cell cycle indicator (FUCCI) and image analysis or flow cytometry to identify and isolate cells in the inner G1 arrested and outer proliferating regions of 3D spheroids.

Tags

Keywords: 3D Tumor SpheroidsMelanomaCell CycleFUCCIImagingFlow CytometryCell PositionTumor MicroenvironmentHypoxiaDrug PenetrationNecrotic CoreProliferationCell-cell Interaction

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