Preparing Free-Floating Slice Cultures from an Adult Human Brain

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.

Transfer the specimen to a Petri dish containing slicing solution. Using fine surgical tools, carefully remove as much of the remaining meninges as possible. Choose the best specimen orientation for producing slices with the p