feature sizes and high aspect ratios for pattern transfer can be obtained. [6][7][8] Typically, such BCP thin films are prepared by spin-coating a dilute BCP solution onto silicon (Si) substrates. However, these as-cast BCP thin films exhibit a locally segregated, disordered morphology. [9][10][11] Although perpendicular lamellar structures can be obtained by various annealing processes, high defectivity in the resultant thin films reduces the potential for industrial applications. Furthermore, although chemical and physical guides can be used to alleviate the thin film defectivity, [6,[12][13][14][15] BCP thin films that can meet the industrial requirements of 0.01 defects cm −2 have yet to be developed.The difficulty in obtaining defect-free BCP thin films can be attributed to a lack of understanding and experimental documentation in the early-stage evolution of ordered structures and defects in perpendicular lamellar thin films. Previous studies on the evolution of ordered structures in thin films have focused on the formation of parallel lamellae with island-hole structures, [16][17][18] and late-stage defect dynamics, [9,[19][20][21][22][23][24][25][26] particularly using low-χ or parallel cylinder-forming BCPs. Capturing the real time evolution of perpendicular lamellae in situ has been difficult as BCP selfassembly tends to occur in shorter time frames than most characterization equipment can analyze the thin films and the fact that parallel structures tend to form due to interfacial selectivity at the substrate and free surfaces. However, by utilizing newer technology such as high-speed atomic force microscopy (AFM) equipped with a heating unit for in situ characterization of the annealing process in a wide temperature window from room temperature to above the glass transition temperature (T g ) of the BCP, the self-assembly behavior of BCP thin films can be documented in real time and space. Additionally, although differences in the surface free energies (SFEs) between the BCP components tend to induce the preferential interfacial segregation of one block, by balancing the surface affinities of both blocks or introducing chemically modifying layers to tune the chemical affinities of the interfaces, the selective segregation of one block can be prevented, The evolution of well-ordered, sub-10 nm, perpendicular lamellae from a disordered state is documented directly using in situ atomic force microscopy, cross-sectional scanning electron microscopy, and grazing-incidence small angle X-ray scattering. Since block copolymer (BCP) self-assembly occurs on extremely fast time scales, directly imaging the growth of perpendicular lamellae remains challenging. The lack of understanding behind early-stage self-assembly and how metastable defects can affect the kinetic behavior near thermodynamic equilibrium prevents the realization of defect-free thin films. By characterizing the evolution of nanostructures using a high-χ, perpendicular lamella-forming BCP, poly(polyhedral oligomeric silsesquioxane)-blockpoly(2,2,2...