This study is an extended investigation on the formation of the first few monolayers of conjugated poly(fluorene)-based polymer films prepared from solution on defined polar and nonpolar surfaces. In particular, a symmetrical A–B–A triblock copolymer consisting of poly(2-alkylaniline) as A blocks and poly(9,9-dialkylfluorene) as B blocks and a poly(9,9-dialkylfluorene) homopolymer is used for this study. The dependence on drying conditions by means of solvent selection, the influence of a subsequent heat treatment, and the influence of the substrate polarity are investigated for ultrathin films as well as the transition from the first monolayers to the bulk polymer. The study is performed with optical absorption and photoluminescence spectroscopy, and atomic force microscopy to obtain complementary information of optical properties and morphological details. We find that ultrathin films (ca. 1–2 nm) prepared on mica from various solvents form highly defined, flat monolayers at the interface without lateral regularities indicating a dipole–dipole interaction between conjugated-polymer segments and mica surface dipoles. This is further confirmed by bathochromic photoluminescence shifts observed for the ultrathin layers compared to the bulk polymer. Complementary experiments on nonpolar surfaces, highly oriented pyrolytic graphite (HOPG), show a total absence of defined flat films supporting the assumption of a dipole–dipole assisted formation on mica. For increased film thickness on mica (5 nm and more) the homopolymer does not form any regular structures or ordered layers on top of the monolayer. In contrast, the triblock copolymer, shorter in length, revealed a tendency to form a less-defined layer-type growth (3–3.5 nm thick) above the monolayer that was of higher order for higher-boiling-point solvents, indicating that the polymers are found in a different conformation. Moreover, one finds that some solvents that show partial immiscibility with the polymer strongly alter the formation of the film. The observations made for the two different types of polymers allow for an assignment of film-formation driving forces to individual polymer segments and allow for the formulation of a growth model that explains the observed results and indicates the importance of appropriate substrate selection for organic electronic/optoelectronic devices.