Huang Jie, Zhang Ling, Shao Ninghui, Zhang Yongqing, Xu Yuyan, Zhou Yihui, Zhang Delong, Zhang Jin, Lee Hyeon Jeong
Zhejiang Polytechnic Institute, Polytechnic Institute, Zhejiang University, Hangzhou 310058, China.
Liangzhu Laboratory, Zhejiang University, Hangzhou 311121, China.
Chem Biomed Imaging. 2024 Oct 4;3(1):15-24. doi: 10.1021/cbmi.4c00055. eCollection 2025 Jan 27.
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 (-), 3D optical sectioning (), 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.
研究胚胎发生对于理解发育生物学和生殖医学至关重要。其过程需要精确的时空调控，其中脂质代谢起着关键作用。然而，由于技术限制，亚细胞水平上脂质种类的空间动态仍不清楚。为应对这一挑战，我们开发了一种基于受激拉曼散射显微镜的高光谱三维成像与分析方法（超三维受激拉曼散射显微镜，hyper-3D SRS），以通过亚微米分辨率（-）、三维光学切片（）和化学键选择性（Ω）成像来定量评估单个胚胎中的脂质分布。使用超三维受激拉曼散射显微镜，可对单个细胞内的单个脂滴（LDs）进行识别和定量。我们的研究结果表明，即使在二细胞阶段，单个胚胎内的脂滴分布也不均匀。值得注意的是，我们还发现随着胚胎发育，脂滴大小与不饱和度之间存在动态关系，这表明早期发育过程中脂质代谢具有多样性。此外，在早衰小鼠模型的卵母细胞中观察到异常脂滴，这表明脂滴可作为评估卵母细胞/胚胎质量的潜在生物标志物。总体而言，我们的结果突出了超三维受激拉曼散射显微镜作为一种研究胚胎中脂质含量、组成和亚细胞分布的非侵入性方法的潜力。该技术为胚胎发育过程中的脂质代谢提供了有价值的见解，并在评估卵母细胞/胚胎质量方面具有临床应用潜力。
