Improving the photoluminescence quantum yield of conjugated polymer films is very important for color-conversion materials and polymer-based photo-electronic devices. We investigated the effect of the molecular weight and conformation of fluorene/MEH−PPV copolymers on the photoluminescence quantum yield of conjugated polymer films. The films cast from high molecular weight copolymers had high quantum yields (61.2 ± 5.0%) and flexible-coil conformation. The quantum yields of low molecular weight copolymers were significantly lower following film formation (26.8 ± 0.05%) and rod-like conformation. Large reduction in the 0−0 emission band was observed following quantum yield reduction, suggesting increased energy transfer rate. The low molecular weight polymers were prone to easy π-π stack structure formation due to their stretched conformation. This structure promoted the energy transfer rate between polymers and thus the energy that comes to quenching sites increased, leading to the reduction in quantum yield. Although the reduction in the quantum yield of conjugated polymer films is generally attributed to aggregate formation, the red-shift in the absorbance and photoluminescence spectra following film formation, which indicate aggregate formation, was small and uncorrelated with the quantum yield.