This work demonstrates the comparative studies of non-radiative resonance energy transfer in bi-polymer nanoparticles based on fluorescent conjugated polymers. For this purpose, poly[(9,9-dihexylfluorene) (PF) as a donor (D) and poly[2-methoxy-5-(2'-ethyl-hexyloxy)-1,4-phenylene vinylene] (MEH-PPV) as an acceptor (A) have been utilized, from which four different bi-polymer nanoparticle systems are designed and synthesized. Both, steady-state fluorescence spectra and time-resolved fluorescence measurements indicate varying energy transfer efficiencies from the host polymer PF to the acceptor polymer MEH-PPV depending on the D-A distances and structural properties of the nanoparticles. The first approach involves the preparation of PF and MEH-PPV nanoparticles separately and mixing them at a certain ratio. In the second approach, first PF and MEH-PPV solutions are mixed prior to nanoparticle formation and then nanoparticles are prepared from the mixture. Third and fourth approaches involve the sequential nanoparticle preparation. In the former, nanoparticles are prepared to have PF as a core and MEH-PPV as a shell. The latter is the reverse of the third in which the core is MEH-PPV and the shell is PF. The highest energy transfer efficiency recorded to be 35% is obtained from the last system, in which a PF layer is sequentially formed on MEH-PPV NPs. © 2010 Optical Society of America.