We synthesized size-controlled，uniformly distributed, and thioglycolic-stabilized CdTe nanocrystals by wet chemistry method. The multilayer nanocrystals were assembled on Si substrate by bifunctional linker molecule and the resonance energy transfer between different sized CdTe nanocrystals embedded in gelatin film was investigated. The results showed that the multilayer nanocrystals were in a good order because the distance between the layers was nearly the same. The distance between different sized nanocrystals was relatively short in the directly dried sample. We can observe the resonance energy transfer by the red shift in its photoluminescence.