Synchrotron Fourier transform-infrared (FT-IR) and Raman microspectroscopy were applied to investigate changes in the molecular architecture of mouse oocytes and demonstrate the overall morphology of the maturing oocyte. Here we show that differences were identified between immature mouse oocytes at the germinal vesicle (GV) and mature metaphase II (MII) stage when using this technology, without the introduction of any extrinsic markers, labels, or dyes. GV mouse oocytes were found to have a small, centrally located lipid deposit and another larger polar deposit of similar composition. MII oocytes have very large, centrally located lipid deposits. Each lipid deposit for both cell types contains an inner and outer lipid environment that differs in composition. To assess interoocyte variability, line scans were recorded across the diameter of the oocytes and compared from three independent trials (GV, n = 91; MII, n = 172), and the data were analyzed with principal component analysis (PCA). The average spectra and PCA loading plots show distinct and reproducible changes in the CH stretching region that can be used as molecular maturation markers. The method paves the way for developing an independent assay to assess oocyte status during maturation providing new insights into lipid distribution at the single cell level.Fourier transform-infrared (FT-IR) microspectroscopy and Raman microspectroscopy (RMS) are powerful techniques for studying the composition of cells and tissues. The spectra obtained provide a unique molecular fingerprint that can be interpreted based on the macromolecular chemistry of the cell or tissue under investigation. Hitherto no FT-IR/Raman imaging or mapping study has been performed on entire oocytes, and there are only two studies that have utilized the FT-IR technique to investigate oocyte components. 1,2 These FT-IR studies explored protein secondary structure of the zona pellucida 1 and the effect of chilling on the oolemma. 2 In an FT-IR/Raman mapping experiment each pixel is essentially a digital element

Cell surface glycoproteins play vital roles in cellular homeostasis and disease. Antibody recognition of glycosylation on different cells and pathogens is critically important for immune surveillance. Conversely, adverse immune reactions resulting from antibody-carbohydrate interactions have been implicated in the development of autoimmune diseases and impact areas such as xenotransplantation and cancer treatment. Understanding the nature of antibody-carbohydrate interactions and the method by which saccharides fit into antibody binding sites is important in understanding the recognition process. In silico techniques offer attractive alternatives to experimental methods (X-ray crystallography and NMR) for the study of antibody-carbohydrate complexes. In particular, molecular docking provides information about protein-ligand interactions in systems that are difficult to study with experimental techniques. Before molecular docking can be used to investigate antibody-carbohydrate complexes, validation of an appropriate docking method is required. In this study, four popular docking programs, Glide, AutoDock, GOLD, and FlexX, were assessed for their ability to accurately dock carbohydrates to antibodies. Comparison of top ranking poses with crystal structures highlighted the strengths and weaknesses of these programs. Rigid docking, in which the protein conformation remains static, and flexible docking, where both the protein and ligand are treated as flexible, were compared. This study has revealed that generally molecular docking of carbohydrates to antibodies has been performed best by Glide.