Live Cell Imaging of Chromosome Segregation During Mitosis

作者： Prajakta Varadkar1, Kazuyo Takeda2, Brenton McCright1 1Division of Cellular and Gene Therapies, Center for Biologics Evaluation and Research,US Food and Drug Administration, 2Microscopy and Imaging Core facility, Division of Viral Products, Center for Biologics Evaluation and Research,US Food and Drug Administration

简介：

Overview

This protocol describes a method to label and visualize live chromosomes in mitotic cells using Histone2B-GFP BacMam 2.0 and spinning disc confocal microscopy. This technique allows researchers to study chromosome segregation in cultured cells.

Key Study Components

Area of Science

• Cell Biology
• Microscopy
• Chromosome Dynamics

Background

• Live imaging of chromosomes is crucial for understanding mitosis.
• Histone2B-GFP provides a fluorescent marker for visualization.
• Spinning disc confocal microscopy enhances imaging quality.
• This method can be applied to various genetic and pharmacological studies.

Purpose of Study

• To visualize live chromosomes during mitosis.
• To assess the impact of genetic modifications on chromosome behavior.
• To evaluate drug effects on chromosome segregation.

Methods Used

• Labeling of chromosomes with Histone2B-GFP BacMam 2.0.
• Use of spinning disc confocal microscopy for imaging.
• Synchronization of mouse embryonic fibroblasts (MEFs) in G0/G1 phase.
• Incubation of MEFs in specific growth medium conditions.

Main Results

• Successful labeling of live chromosomes in mitotic MEFs.
• Clear visualization of chromosome dynamics during cell division.
• Demonstration of the method's ease and convenience.
• Potential applications in genetic and drug treatment studies.

Conclusions

• This method provides a reliable approach for live chromosome imaging.
• It facilitates the study of chromosome behavior under various conditions.
• The technique is accessible for researchers in cell biology.

Frequently Asked Questions