Preparing Free-Floating Slice Cultures from an Adult Human Brain
Transcription
Take a human brain tissue immersed in a slicing solution. The solution's biological pH and nutrients help maintain tissue functionality.
Remove the outermost meninges layer, and trim the tissue to create a flat surface.
Blot excess solution from the tissue. Attach the trimmed tissue to a specimen disk with an agarose block, keeping the tissue in contact with the block for ease of sectioning.
Place the disk in a vibratome buffer tray and cover the tissue with an ice-cold slicing solution.
Using the vibratome, generate uniform slices that preserve the tissue architecture.
Trim off excess tissue, ensuring a consistent proportion of gray and white matter.
Place the slices in a multiwell plate containing a culture medium and incubate.
The slices remain suspended in the medium to ensure even nutrient access, a technique known as free-floating slice culture.
Regularly replenish the culture with a neural growth factor-supplemented medium to maintain tissue viability.
