The mechanical properties of laser powder bed fused (LPBFed) Invar 36 alloy have been limited by the presence of manufacturing defects. It is imperative to investigate the influence of these defects on the mechanical behavior of LPBFed Invar 36 alloy. In this study, in-situ X-ray computed tomography (XCT) tests were conducted on LPBFed Invar 36 alloy fabricated at different scanning speeds to examine the relationship between manufacturing defects and mechanical behavior. For LPBFed Invar 36 alloy fabricated at a scanning speed of 400 mm/s, the manufacturing defects were randomly distributed and tended to be elliptical in shape. Plastic deformation behavior was observed, and failure initiated from defects inside the material resulting in ductile failure. Conversely, for LPBFed Invar 36 alloy fabricated at a scanning speed of 1000 mm/s, numerous lamellar defects were observed mainly located between deposition layers, and their quantity was significantly increased. Little plastic deformation behavior was observed, and failure initiated from defects on the shallow surface of the material resulting in brittle failure. The differences in manufacturing defects and mechanical behavior are attributed to changes in input energy during the laser powder bed fusion process.