Recent insights suggest that the surrounding environment of the pre-implantation embryo is likely to alter long-term trajectory. The periconception environment therefore represents a critical window for programming fetal growth. This study evaluated the impact of the two major clinical components of assisted reproductive technology (ART), embryo culture and ovarian hyperstimulation, on embryo development by using three metabolic markers: Peroxyfluor 1 (PF1), to assess hydrogen peroxide levels, Monochlorobimane (MCB), to assess reduced glutathione abundance and Mitotracker Deep Red, to detect active mitochondria. In addition to assessment of the embryos based on morphology and staining intensity, textural analysis using Grey Level Co-occurrence Matrix (GLCM), a second-order statistical model was used to evaluate PF1, MCB and MTDR staining, providing a robust metric of embryo health. Embryos were collected 48, 60 and 88 h post-hCG treatment, corresponding to the 2-cell, 8-cell and blastocyst stage. Our results showed that embryos derived from ovarian hyperstimulation had significantly higher intensity and texture heterogeneity of active mitochondria and hydrogen peroxide as compared to the natural cycling embryos. Embryos exposed to embryo culture displayed variations in texture of active mitochondria, although there was no change in intensity. Our data provide strong evidence that the metabolic profiling and texture were modified in embryos derived from ART. To the best of our knowledge, this is one of the first studies to investigate the metabolic profiling using texture analysis in embryos, although the functional roles of each texture features are not yet well understood. This study supports that textural analysis provides a means of gaining additional information regarding sub-cellular analyses instead of using intensity measurements alone.