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Full field optical coherence tomography can identify spermatogenesis in a rodent sertoli-cell only model

机译:全场光学相干断层扫描可以识别仅在啮齿动物的睾丸干细胞模型中的精子发生

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Background:Microdissection testicular sperm extraction (micro-TESE) has replaced conventional testis biopsies as a method of choice for obtaining sperm for in vitro fertilization for men with nonobstructive azoospermia. A technical challenge of micro-TESE is that the low magnification inspection of the tubules with a surgical microscope is insufficient to definitively identify sperm-containing tubules, necessitating tissue removal and cytologic assessment. Full field optical coherence tomography (FFOCT) uses white light interference microscopy to generate quick high-resolution tomographic images of fresh (unprocessed and unstained) tissue. Furthermore, by using a nonlaser safe light source (150 W halogen lamp) for tissue illumination, it ensures that the sperm extracted for in vitro fertilization are not photo-damaged or mutagenized.Materials and Methods:A focal Sertoli-cell only rodent model was created with busulfan injection in adult rats. Ex vivo testicular tissues from both normal and busulfan-treated rats were imaged with a commercial modified FFOCT system, Light-CT?, and the images were correlated with gold standard hematoxylin and eosin staining.Results:Light-CT? identified spermatogenesis within the seminiferous tubules in freshly excised testicular tissue, without the use of exogenous contrast or fixation. Normal adult rats exhibited tubules with uniform size and shape (diameter 328 ±11 μm). The busulfan-treated animals showed marked heterogeneity in tubular size and shape (diameter 178 ± 35 μm) and only 10% contained sperm within the lumen.Conclusion:FFOCT has the potential to facilitate real-time visualization of spermatogenesis in humans, and aid in micro-TESE for men with infertility.
机译:背景:显微解剖睾丸精子提取术(micro-TESE)代替了常规的睾丸活检术,作为获得非阻塞性无精子症男性精子以进行体外受精的一种选择方法。 micro-TESE的一项技术挑战是,用手术显微镜对肾小管进行低倍检查不足以明确确定含精子的肾小管,因此需要进行组织切除和细胞学评估。全场光学相干断层扫描(FFOCT)使用白光干涉显微镜来生成新鲜(未处理和未染色)组织的快速高分辨率断层图像。此外,通过使用非激光安全光源(150 W卤素灯)进行组织照明,可以确保提取的用于体外受精的精子不会受到光致损伤或诱变。材料与方法:建立了仅局灶性支持细胞的啮齿动物模型用白消安注射液在成年大鼠中产生。使用商业改良的FFOCT系统Light-CT?对正常和白消安治疗的大鼠的离体睾丸组织成像,并将这些图像与金标准苏木精和曙红染色相关。无需使用外源性对比剂或固定剂,即可在刚切除的睾丸组织中的生精小管内鉴定出精子发生。正常成年大鼠的肾小管的大小和形状均一(直径328±11μm)。经环丁砜处理的动物的肾小管大小和形状(直径178±35μm)表现出明显的异质性,腔内精子中仅含10%的精子。结论:FFOCT有潜力促进人类精子发生的实时可视化,并有助于micro-TESE用于不育男性。

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