Abstract
Time-of-flight secondary ion mass spectrometry (ToF-SIMS) imaging has been used for the direct analysis of single intact Xenopus laevis embryo surfaces, locating multiple lipids during fertilization and the early embryo development stages with subcellular lateral resolution (∼4 μm). The method avoids the complicated sample preparation for lipid analysis of the embryos, which requires selective chemical extraction of a pool of samples and chromatographic separation, while preserving the spatial distribution of biological species. The results show ToF-SIMS is capable of profiling multiple components (e.g., glycerophosphocholine, SM, cholesterol, vitamin E, diacylglycerol, and triacylglycerol) in a single X. laevis embryo. We observe lipid remodeling during fertilization and early embryo development via time course sampling. The study also reveals the lipid distribution on the gamete fusion site. The methodology used in the study opens the possibility of studying developmental biology using high resolution imaging MS and of understanding the functional role of the biological molecules.
Highlights
Time-of-flight secondary ion mass spectrometry (ToF-SIMS) imaging has been used for the direct analysis of single intact Xenopus laevis embryo surfaces, locating multiple lipids during fertilization and the early embryo development stages with subcellular lateral resolution (ف4 m)
Techniques such as scanning electron microscopy and confocal microscopy have been applied to image the frog egg and embryo to investigate the mechanisms of biological processes and embryo development
Most synthesized GPCho is transported away from the center to form the outer layer of the fusion site
Summary
Time-of-flight secondary ion mass spectrometry (ToF-SIMS) imaging has been used for the direct analysis of single intact Xenopus laevis embryo surfaces, locating multiple lipids during fertilization and the early embryo development stages with subcellular lateral resolution (ف4 m). Spatiotemporal lipid profiling during early embryo development of Xenopus laevis using dynamic ToF-SIMS imaging. The detected compounds include organic acids, fatty acids (saturated, unsaturated, and hydroxy fatty acids ranging from C8 to C28), amino acids, alcohols, and sugars These techniques, based on solvent extraction, do not provide any information on spatial localization beyond the fraction which was extracted; they offer qualitative and quantitative information about the composition of the cytoplasm with respect to key lipid compounds [6]. A current challenge in lipid biology studies is determining the spatial location of multiple lipids while retaining compositional information Techniques such as scanning electron microscopy and confocal microscopy have been applied to image the frog egg and embryo to investigate the mechanisms of biological processes and embryo development. The architecture of the membrane exhibits dramatic changes before and after
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