The main objective of the proposal is to develop a new easy-to-use tool for in-vitro screening of neurotoxic agents, based on changes/disappearance of electrophysiological activity in acute brain slices and organotypic slice cultures as a consequence of their exposure to neurotoxic agents. The proposal will take advantage of silicon technology by developing a micromachined multi-electrode array to be functionally coupled to brain slices and organotypic slice cultures. Thus allowing multi-site measurements repeatable in time.
The ease of use of the new tool should rely on the design of an optimised measurement micro-chamber for acute brain slices and organotypic brain slices. The core of the microchambe1 will be the array of micro-electrode on which biological samples will lay. This recording /stimulating array will form the bottom of the micro-chamber. The micro-chamber should guarantee the best possible approximation to physiological conditions (Cf. "Central Nervous System Slice Preparations", 3rd Int. Conf., June 1994, J. of Neurosci. Methods, 59, 1, 1995).
Appropriate software / hardware tools will be developed to optimise multi-site recording/stimulation of the electrophysiological signals and their processing in order to "translate" them into a highly reproducible neurotoxicology screening test. The development and use of the new array - based micro-chamber will be paralleled during the proposal by histological, biochemical and standard electrophysiology tests.
Measurable objectives to be achieved in the design of the chambers are:
1) biocompatibility of all the materials in contact with the live neural cell cultures,
2) chemical stability of chosen materials in saline and culture media, 3) automated multi-site recording and stimulation and
4) ease of handling.
The global measurable objective is to provide the European Community with a new tool for in-vitro electrophysiological tests which will reduce animal experiments and become an improved alternative to other time-consuming and expensive in-vitro experiments.
The performances of the micro-chamber will be validated by testing a set of known neurotoxic agents.
Funding SchemeCSC - Cost-sharing contracts