Lipid Metabolic Heterogeneity during Early Embryogenesis Revealed by Hyper-3D Stimulated Raman Imaging

Studying embryogenesis is fundamental to understanding developmental biology and reproductive medicine. Its process requires precise spatiotemporal regulations in which lipid metabolism plays a crucial role. However, the spatial dynamics of lipid species at the subcellular level remains obscure due to technical limitations. To address this challenge, we developed a hyperspectral 3D imaging and analysis method based on stimulated Raman scattering microscopy (hyper-3D SRS) to quantitatively assess lipid profiles in individual embryos through submicrometer resolution (x-y), 3D optical sectioning (z), and chemical bond-selective (Ω) imaging. Using hyper-3D SRS, individual lipid droplets (LDs) in single cells were identified and quantified. Our findings revealed that the LD profiles within a single embryo are not uniform, even as early as the 2-cell stage. Notably, we also discovered a dynamic relationship between the LD size and unsaturation degree as embryos develop, indicating diverse lipid metabolism during early development. Furthermore, abnormal LDs were observed in oocytes of a progeria mouse model, suggesting that LDs could serve as a potential biomarker for assessing oocyte/embryo quality. Overall, our results highlight the potential of hyper-3D SRS as a noninvasive method for studying lipid content, composition, and subcellular distribution in embryos. This technique provides valuable insights into lipid metabolism during embryonic development and has the potential for clinical applications in evaluating oocyte/embryo quality.