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Multi-color Localization Microscopy of Single Membrane Proteins in Organelles of Live Mammalian Cells
Published on: June 30, 2018
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Analysis of the Localization of MEN Components by Live Cell Imaging Microscopy.
1Department of Genetics, University of Cambridge, Downing Street, Cambridge, CB2 3EH, UK.
Methods in Molecular Biology (Clifton, N.J.)
|November 10, 2016
Summary
This study details yeast live cell microscopy protocols to understand how cells decide when to exit mitosis. These methods help visualize the mitotic exit network (MEN) pathway, crucial for cell division.
Area of Science:
- Cell Biology
- Molecular Biology
- Genetics
Background:
- Mitotic exit is a critical cell division process regulated by spatial and temporal cues.
- The mitotic exit network (MEN) integrates these signals to control cell cycle progression.
- Understanding MEN regulation is key to comprehending cell division fidelity.
Purpose of the Study:
- To present robust protocols for yeast live cell microscopy.
- To facilitate the dissection of the mitotic exit decision-making process.
- To enable visualization of the mitotic exit network (MEN) pathway dynamics.
Main Methods:
- Live cell imaging of budding yeast.
- Utilizing fluorescent protein tags for protein localization and dynamics.
- Standardized microscopy protocols for reproducible results.
Main Results:
- Established reliable live cell microscopy techniques for yeast.
- Demonstrated the applicability of these methods to study MEN signaling.
- Provided a foundation for dissecting spatial-temporal regulation of mitotic exit.
Conclusions:
- Routine yeast live cell microscopy is effective for studying complex cell cycle events.
- The presented protocols aid in the investigation of the mitotic exit network (MEN).
- This work supports further research into the spatial and temporal control of cell division.
Keywords:
Cell population analysisDigital imageFluorescence microscopyFluorescent protein tagsLive cell imagingSingle cell analysisStill imagingTime lapseMore Related Videos
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