The experiments described here were designed to characterize the location and orientation of a single homopolymer chain embedded in a block copolymer lamella. We used scanning near-field optical microscopy (SNOM) to image individual perylene-labeled poly(methyl methacrylate) (PMMA-Pe) chains sparsely distributed in lamellar phases formed by poly(styrene-b-methyl methacrylate) (PS-PMMA). By calculating the center of mass (CM) of the PMMA-Pe chain, we found that the homopolymer chains were distributed throughout the PMMA-rich domain layers, with the maximum CM population at the domain centers. The orientation of the homopolymer chains is dependent on the location of the CM of the homopolymer in the PMMA block domain. If the CM locates at the domain center, the homopolymer chain prefers an orientation parallel with the lamella. If the CM is close to the block interface, the homopolymer chain tends to orient itself perpendicularly to the interface. This location dependence of chain orientation is likely the result of the rotational freedom of PMMA-Pe chains in the PMMA-rich domains of the PS-PMMA block copolymer lamella.