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Combining Mitotic Cell Synchronization and High Resolution Confocal Microscopy to Study the Role of Multifunctional Cell Cycle Proteins During Mitosis
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Biologia
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JoVE Journal Biologia
Combining Mitotic Cell Synchronization and High Resolution Confocal Microscopy to Study the Role of Multifunctional Cell Cycle Proteins During Mitosis

Combining Mitotic Cell Synchronization and High Resolution Confocal Microscopy to Study the Role of Multifunctional Cell Cycle Proteins During Mitosis

DOI:
10.3791/56513-v

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08:33 min

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December 05, 2017

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1Department of Cell and Molecular Biology, Feinberg School of Medicine,Northwestern University

Capitoli

  • 00:05Titolo
  • 00:52Mitotic Cell Progression Fixed Cell Imaging
  • 03:35Mitotic Cell Progression Live Cell Imaging
  • 05:31Results: Representative Mitotic Progression in Control- and siRNA-depleted Cells
  • 07:09Conclusion

Summary

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We present a protocol for double thymidine synchronization of HeLa cells followed by analysis using high resolution confocal microscopy. This method is key to obtaining large number of cells that proceed synchronously from S phase to mitosis, enabling studies on mitotic roles of multifunctional proteins which also possess interphase functions.

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Keywords: Cell CycleMitosisCell SynchronizationConfocal MicroscopyMultifunctional ProteinsHeLa CellsThymidine BlockSiRNAImmunofluorescence

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