We investigate the role of the chain length and molecular weight distribution on the diffusion dynamics of freshly synthesized MEH-PPV polymer chains, For the above purpose, a new technique based on combination of size exclusion chromatography (SEC) with fluorescence correlation spectroscopy (FCS) is developed to probe the diffusion dynamics of a narrow molecular weight distribution of fractionated samples of 20-500 kDa. The narrow dispersed samples were characterized by absorbance, emission, and time-resolved fluorescence decay techniques. The results revealed that the properties of fractionated samples were almost uniform for a wide range of molecular weights. A maximum entropy based method for FCS data analysis is employed to obtain the correct diffusion coefficients of the polymer chains with heterogeneous dynamics. The FCS experiment on the unfractionated broad molecular weight sample is not enough to establish the correlation between the molecular weight of the chains with diffusion dynamics and emphasized the need for relatively monodispersed pi-conjugated polymers. FCS results show that higher molecular weight chains diffuse much faster than shorter ones. Atomic force microscopy revealed that 300 kDa polymers produced 130 nm particles, whereas 50 kDa polymer chains formed micrometer size aggregates. At higher molecular weights, the strong chain interactions promote the formation of globular (or tightly packed) particles which diffuse faster in solution. The low molecular weight chains experience strong interparticle interaction; as a consequence, the diffusion of chains becomes slower. In the present investigation, we demonstrate the need for the narrow polydisperse sample for establishing the correlation between diffusion dynamics and chain length (or molecular weights) of pi-conjugated polymers using a single molecule spectroscopy technique such as FCS.