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Analysis of Mouse Brain Sections by Live-cell Time-lapse Confocal Microscopy
Tao Yang1,2,3, Ty Hergenreder1, Bing Ye1,2
1Life Sciences Institute, University of Michigan, Ann Arbor, MI 48109, USA.
Bio-Protocol
|April 14, 2023
Summary
Researchers developed a novel imaging technique to observe neuron migration in the developing brain. This method allows for detailed study of cellular events crucial for proper brain development and may aid in understanding neurological disorders.
Area of Science:
- Neuroscience
- Developmental Biology
- Cell Biology
Background:
- Mammalian cerebral cortex development relies on radial migration of pyramidal neurons from the neural tube.
- Aberrations in neuron migration can lead to structural changes and neurological disorders.
- Understanding neuron migration dynamics requires high-resolution, long-term imaging.
Purpose of the Study:
- To develop and share a protocol for observing cellular events during mammalian pyramidal neuron migration.
- To enable high-resolution, time-lapse imaging of neuron migration over extended periods.
- To provide a tool for studying neurodevelopmental and pathological processes.
Main Methods:
- Established an organotypic brain slice preparation compatible with confocal microscopy.
- Utilized a Leica SP5 confocal microscope for time-lapse imaging over 12 hours.
- Developed guidelines for overcoming technical challenges in setting up the imaging system.
Main Results:
- Successfully implemented a system for prolonged, high-resolution imaging of neuron migration.
- Demonstrated the feasibility of observing cellular events during radial migration in brain slices.
- The developed protocol facilitates detailed study of neuron positioning and its impact on cortical thickness.
Conclusions:
- The presented protocol enables unprecedented observation of neuron migration dynamics.
- This technique is valuable for investigating normal neurodevelopment and neurological disorders.
- The system supports the study of various cellular events beyond neuron migration.
