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超薄冷冻切片的相关显微镜检查是胎盘研究的有力工具。

Correlative microscopy of ultrathin cryosections is a powerful tool for placental research.

作者信息

Takizawa T, Robinson J M

机构信息

Department of Physiology and Cell Biology, Ohio State University, Columbus, OH 43210, USA.

出版信息

Placenta. 2003 May;24(5):557-65. doi: 10.1053/plac.2002.0942.

Abstract

In this report, we describe procedures for correlative fluorescence and electron microscopy in immunocytochemical studies on the human placenta. Ultrathin cryosections of placenta were used for detection of the distribution of antigens by immunofluorescence and subsequently by immunoelectron microscopy of the same ultrathin cryosection. This methodology has certain advantages over conventional immunohistochemistry and immunoelectron microscopy. The advantages are, most notably, that the same exact structures are examined by both imaging modalities. In addition, since the tissue is physically sectioned (50-100 nm thickness), greater resolution for fluorescence can be obtained in the z-dimension than can be obtained by optical sectioning in confocal microscopy. This last point is of particular importance for discriminating between structures closely stacked in the z-dimension. In this report, we have determined the distribution of caveolin-1 in ultrathin cryosections of terminal villi of the human term placenta. We demonstrate that the use of ultrathin cryosections is a powerful approach for immunofluorescence and correlative microscopy for the in situ localization of antigens.

摘要

在本报告中,我们描述了在人胎盘免疫细胞化学研究中进行相关荧光和电子显微镜检查的方法。胎盘超薄冰冻切片用于通过免疫荧光检测抗原分布,随后对同一超薄冰冻切片进行免疫电子显微镜检查。该方法相对于传统免疫组织化学和免疫电子显微镜具有某些优势。最显著的优势是,两种成像方式检查的是完全相同的结构。此外,由于组织是进行物理切片(厚度为50 - 100纳米),与共聚焦显微镜的光学切片相比,在z维度上荧光可获得更高的分辨率。最后这一点对于区分在z维度上紧密堆叠的结构尤为重要。在本报告中,我们确定了人足月胎盘终末绒毛超薄冰冻切片中小窝蛋白-1的分布。我们证明,使用超薄冰冻切片是一种用于免疫荧光和相关显微镜检查以进行抗原原位定位的强大方法。

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