The structure evolution of iPS during isothermal cold-crystallization and subsequent melting processes is studied by in situ monitoring its FTIR spectra. Through following the intensity changes of the characteristic bands associated with its crystalline status and 3 1 helix conformation chains, the multiple melting behavior of iPS cold-crystallized at different temperatures has been discussed in detail. It is concluded that (i) the lowest-temperature annealing peak can be ascribed either to the relaxing of locally ordered long 3 1 helix chains in the amorphous phase or to the melting of some microcrystallites formed upon long-time annealing; (ii) the double melting behavior of iPS cold-crystallized at temperatures lower than 180°C is mainly attributed to the model of melting, recrystallization, and remelting of the iPS crystals; and (iii) for the sample cold-crystallized at a temperature of 200°C, the in situ IR experiments give strong evidence to support the existence of different kinds of crystals which show different thermodynamic stabilities.