N,N′-bis(9H-fluoren-9-ylidene)benzene-1,4-diamine was synthesized via the acetic acid-assisted Schiff base reaction between 9-fluorenone and p-phenylenediamine. The thin films were deposited from solution and characterized by contact angle measurements (CAM), X-ray photoelectron spectroscopy (XPS) and tunneling conductance spectroscopy (TCS). The tunneling conductance spectra, related to the potential and distance between the tip and substrate, were acquired at different tip–substrate separations and depicted significant trend under the action of electric field. Systematic analysis shows more information about electron transport through medium layers. The electric field plays an important role in tunneling conductance spectra. The tunneling conductance spectra data indicate the electric field dependence of electron transport.