https://scholars.lib.ntu.edu.tw/handle/123456789/460299
標題: | Higher harmonie generation microscopy of in vitro cultured mammal oocytes and embryos | 作者: | Hsieh C.-S SHEE-UAN CHEN Lee Y.-W YU-SHIH YANG CHI-KUANG SUN |
公開日期: | 2008 | 卷: | 16 | 期: | 15 | 起(迄)頁: | 11574-11588 | 來源出版物: | Optics Express | 摘要: | Oocyte and embryo selection governs the success of assisted reproductive technologies. The imaging tools applied for selecting embryos may need to contain several key properties: noninvasiveness, high 3D resolution, and the contrast capability to provide as much information about the embryos as possible, such as spindle fibers, zona pellucida, and organelles. Currently adopted imaging techniques can only provide one or two of these desired properties and are with limited contrast of the embryos. Some image techniques can even damage the embryos. Previous studies have shown that harmonic generation microscopy (HGM), a virtual-transition based technology, can provide noninvasive imaging in zebrafish embryos with a sub-cellular 3D resolution and a millimeter penetration depth, and thus could be a suitable tool for future oocyte and embryo selection of assisted reproductive technologies. However to evaluate HGM in clinical use, the intrinsic contrast origin of the second harmonic generation (SHG) and third harmonic generation (THG) inside the mammal embryos has to be studied. In this work we performed HGM studies on the in vitro cultured mouse oocytes and embryos by combining the SHG and THG modalities, with a focus on the contrast origin evaluation. Through the noninvasive HGM imaging, we can clearly identify various structures in the whole oocytes and embryos, including spindle fibers, zona pellucida, polar bodies, cell membranes, and the laminated organelles in the cells. The origin of the THG contrast was further confirmed through the standard staining studies. Through SHG signals, we could not only observe the spindle fibers when the oocytes were arrested at metaphase II or during the cleavage of the embryos, but can also distinguish and analyze the thickness of the three layers of the zona pellucida. Combining two different higher-harmonic generation modalities, SHG and THG, HGM successfully revealed the subcellular structures of the whole mouse embryos with a high 3D spatial resolution. ? 2008 Optical Society of America. |
URI: | https://scholars.lib.ntu.edu.tw/handle/123456789/460299 | ISSN: | 1094-4087 | SDG/關鍵字: | Cytology; Harmonic analysis; Imaging techniques; Mammals; Nonlinear optics; Assisted reproductive technologies; Harmonic generation microscopy; Non-invasive imaging; Spatial resolution; Subcellular structure; Virtual transitions; Zebrafish embryos; Zona pellucida; Harmonic generation; animal; article; cell culture; confocal microscopy; cytology; embryo (anatomy); female; fluorescence microscopy; image enhancement; Institute for Cancer Research mouse; methodology; mouse; oocyte; organ culture technique; Animals; Cells, Cultured; Embryo, Mammalian; Female; Image Enhancement; Mice; Mice, Inbred ICR; Microscopy, Confocal; Microscopy, Fluorescence; Oocytes; Organ Culture Techniques |
顯示於: | 醫學系 |
在 IR 系統中的文件,除了特別指名其著作權條款之外,均受到著作權保護,並且保留所有的權利。