A method for defined sectioning of fresh young brains and collection of small regions for cell and tissue culture.

A method for the collection of defined small regions from fresh brain under sterile conditions is described. Its reproducibility allows the cultivation of well-defined, corresponding regions from similar brains. After controlled orientation in agarose, the brain can be sectioned by means of a Vibratome in spite of its softness. The section level is defined by co-ordinates of a stereotaxic atlas, produced for this purpose from brains orientated in the same way. Areas desired for cultivation are punched out from tissue slices with 200-550 microns diameter needles according to the atlas pictures. The plugs can then be stored in cold buffer solution until preparation for culture. Exact locations of tissue samples collected can be determined histologically. Either whole or dissociated plugs cultivated by a plasma clot technique lead to morphologically differentiating neurons surviving for more than or up to 14 days, respectively.