The microstructure of aluminum substrates was investigated during the quality control of grain sizes determination and crystal orientation analysis before and after preheat (recrystallization) treatment at 500 C, in relation to the microstructure and heat stability of nanoporous anodic alumina (NPAA) membranes formed on various preheated aluminum substrates and subsequently detached. Depending on the preheating conditions, the average grain size of the aluminum substrate increased from 3 mm to 1280 mm. Also, the preferential crystal orientation of aluminum was (311) and/or (100) or (311)/(422) or (220) under different heating conditions. Recrystallization at a higher heating rate and particularly a higher cooling rate yielded lower grain growth of the aluminum substrate, so that quenching rather than annealing resulted in smaller grains and domains as well as a larger number of grain boundaries on the top surface of related a-NPAA membranes. This induced less ordered growth in the nanopore structure and lower stability against cracking and deflection during crystallizing a-NPAA thin film membranes. Figure 3. Optical microscopy images of detached as-anodized NPAA film formed on differently treated Al substrates: (a) NPAA-As-polished, (b) NPAA-IQ, (c) NPAA-LL1, and (d) NPAA-LL3.a-Porous anodic alumina with pore regularity and thermal stability