Photoresist (PR) profiles tend to have deformation and shrinkage with typical transmission electron microscopy (TEM) sample preparation methods using a focused ion beam scanning electron microscope (FIB-SEM). As the temperature increases during the TEM sample preparation, it may lead to deformation and shrinkage in PR profiles. In this study, we analyze the impact when performing the sample preparation at a cold temperature using a cryo-FIB to minimize deformation and shrinkage issues. To test this methodology, the TEM sample preparation process was performed under different conditions. From these experiments, the TEM results with full cryo conditions showed that the PR line to space ratio was closest to the target, which is the sample’s real line to space ratio (1:1), and the bottom anti-reflective coating (BARC) shrinkage was minimized.