LB films of a novel azobenzene derivative were fabricated and characterized by AFM, Raman microscopy, and polarized transmission FTIR spectroscopy. The AFM and Raman microscopic data revealed a well-ordered uniform structure of the film, and the dichroism in two perpendicular polarized FTIR suggested that there exist two types of hydrogen-bonded carboxyl groups, which are orientated in planes perpendicular to each other, forming a two-dimensional inter-molecular hydrogen-bonded structure in the LB film.