Objective This study aims to identify the change in anthropometric measurements during spaceflight due to microgravity exposure. Background Comprehensive and accurate anthropometric measurements are crucial to assess body shape and size changes in microgravity. However, only limited anthropometric data have been available from the astronauts in spaceflight. Methods A new photogrammetry-based technique in combination with a tape-measure method was used for anthropometric measurements from nine crewmembers on the International Space Station. Measurements included circumference and height for body segments (chest, waist, bicep, thigh, calf). The time-dependent variations were also assessed across pre-, in-, and postflight conditions. Results Stature showed a biphasic change with up to 3% increase at the early flight phase, followed by a steady phase during the remaining flight. Postflight measurements returned to a similar level of the preflight. Other linear measurements, including acromion height, showed similar trends. The chest, hip, thigh, and calf circumferences show overall decrease during the flight up to 11%, then returned close to the preflight measurement at postflight. Conclusion The measurements from this study provide critical information for the spacesuit and hardware design. The ground-based assessments for spacesuit fit needs to be revalidated and adjusted for in-flight extravehicular activities from this data. Application These data can be useful for space suit design as well as habitat, vehicle, and additional microgravity activities such as exercise, where the body shape changes can affect fit, performance, and human factors of the overall design.