The first method is extremely labor intensive, requires highly specialized and trained personnel and costly equipment. The success rate for attaching the slice to membrane is low. The second procedure precludes replacing a slice back into its culture conditions once the slice has been removed for a given experiment; chances for attachment of the slice to the membrane before 5 days in culture are very low. Furthermore, coating of the membrane is problematic, and autoclaving is not possible. If individual slices are to be used for experimentation, the membranes can be used only once per slice and cost ~$5. The high cost of the membranes, together with the low likelihood of initial slice attachment make this procedure considerably expensive.

New and improved method.   We have combined the advantages of these two successful methods. We devised a novel tissue-friendly porous surface that brain slices will readily adhere to as soon as the first day in culture with a remarkably high success rate. The absence of clot, or any other foreign substance, to attach the cultures to their supporting membrane should greatly facilitate studies of the synaptic reorganization and sprouting that occur during the first days in culture and should be helpful for studying the mechanisms implicated in developmental neuronal plasticity.

If necessary, this filter can be easily coated with different substances. After coating, the coverslip and the attached membrane can be autoclaved if needed, a procedure that cannot be performed using the other culture methods. The attachment of the slice to the supporting coverslip also allows for easy manipulation of the slices without disturbing their physical structure. This includes removal of the slices from the culture environment, followed by some experimentation (e.g., electrophysiological or biochemical), and subsequent replacement of the slice together with its coverslip into the culture environment.

Our novel approach is very simple, no specific product or handling is required, no plasma clot or other substances are necessary for attachment, and no roller drum has to be used. This procedure is extremely economical, and thus has the potential to revolutionize medical and biological experimentation in organotypic slice cultures.

Summary of the advantages over previous techniques: