In this work,the single-chain elasticity of polyformaldehyde(POM)is studied,for the first time,by employing atomic force microscopy(AFM)-based single molecule force spectroscopy(SMFS).We find that the single-chain elasticity of POM in a nonpolar organic solvent(nonane)can be described well by a theoretical model(QM-FRC model),when the rotating unit length is 0.144 nm(C-O bond length).After comparison,POM is more flexible than polystyrene(a typical polymer with C-C backbone)at the single-chain level,which is reasonable since the C-O bond has a lower rotation barrier than C-C bond.This result indicates that the flexibility of a polymer chain can be tuned by the C-O bond proportion in backbone,which casts new light on the rational design of new synthetic polymers in the future.