Absorption and fluorescence spectra of 4-cyano-4’-n-octyloxybiphenyl (80CB) films deposited on hydrophilic and hydrophobic substrates were measured and compared with those of its neat samples (crystalline and liquid crystal states). 80CB films deposited on hydrophilic substrates were crystalline and showed monomer-like fluorescence, while 80CB Alms on hydrophobic substrates, where hydrophilic glass sites for 80CB molecules to attach to were absent, were amorphous, showed excimer fluorescence, and had the similar absorption spectrum to that in solution. The deposition processes of 80CB were investigated by measuring in-situ fluorescence spectra during deposition on hydrophilic and hydrophobic substrates. A discontinuous change was found in the series of fluorescence spectra observed during deposition on hydrophilic substrates, while such a change was not found for deposition on the above-hydrophobic substrates. The change was ascribed to structural transformation from amorphous to crystalline states. The average film thickness where the transformation occurred depended on the nature of the substrate surface and the deposition temperature. The difference in the fluorescence behavior between hydrophilic and hydrophobic substrates was discussed in terms of molecular orientation of 80CB in the monolayer on substrates.