Abstract: A chip and an apparatus for acetylcholinesterase inhibitor detection and a method for using thereof are provided. The chip of the invention comprises a substrate and a film, coated on said substrate, including a molecule with a specific ability to combine with an acetylcholinesterase inhibitor. The present invention utilizes the surface modification technology and cooperates with optical precision instruments to provide an acetylcholineases inhibitor detection chip and an apparatus with high sensitivity and suitable for both liquid and gas detection.