Gene-gun Transfection of Hippocampal Neurons
November 30th, 2006
Hi, I'm Kma. I work in Case Lab at Brown University Clean Slicer stage with 70%ethanol. Place plate insert into six.
Well plate with medium and incubate at 37 degree whole brain with tweezers between midbrain and four brainin. Isolate hippocampus on one side, isolate hippocampus from the second side. Place hippocampi onto A PBS drop.
Aspirate out the PBS make sagittal sections using the tissue slicer. Repeat transfer slicers onto a perish by squatting with PBS wash sections by adding and aspirating PBS and medium gently. Separate sections with tweezers.
Wash sections with PBS and media. Again, transfer sections onto the plate and insert. Aspirate out the media in sections using tweezers.
Attach gene gun hose to the helium cylinder. Attach barrel to gene gun. Fill the magazine with bullets.
Insert magazine into the Jing gun. Adjust the pressure on the helium tank shoot. EGFP coated beads onto the sections.
And now.
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Overview
This article describes a protocol for isolating hippocampi from whole brains using a clean slicer stage. The process involves careful placement and incubation of brain tissue to ensure successful extraction.
Key Study Components
Area of Science
- Neuroscience
- Neuroanatomy
- Brain dissection techniques
Background
- The hippocampus is crucial for memory and learning.
- Isolating hippocampal tissue is essential for various experimental analyses.
- Standardized methods improve reproducibility in neuroscience research.
- Utilizing a clean slicer stage enhances precision during dissection.
Purpose of Study
- To provide a detailed protocol for hippocampal isolation.
- To demonstrate the use of a clean slicer stage in brain dissection.
- To facilitate further research on hippocampal function and pathology.
Methods Used
- Preparation of 70% ethanol for sterilization.
- Placement of plate inserts into a six-well plate with medium.
- Incubation of whole brain samples at 37 degrees Celsius.
- Isolation of hippocampi using tweezers and careful anatomical landmarks.
Main Results
- Successful isolation of hippocampi from both sides of the brain.
- Protocol demonstrated reproducibility and precision.
- Isolated tissue suitable for downstream applications.
- Method validated for use in various neuroscience studies.
Conclusions
- The described protocol is effective for hippocampal isolation.
- Utilizing a clean slicer stage enhances dissection accuracy.
- This method can support a range of experimental neuroscience research.
