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用于免疫细胞化学的固定和冷冻保护生物材料的冷冻切片制作

Cryosectioning fixed and cryoprotected biological material for immunocytochemistry.

作者信息

Webster Paul, Webster Alexandre

机构信息

Ahmanson Advanced Electron Microscopy and Imaging Center, House Ear Institute, Los Angeles, CA, USA.

出版信息

Methods Mol Biol. 2007;369:257-89. doi: 10.1007/978-1-59745-294-6_13.

Abstract

Immunocytochemistry for transmission electron microscopy provides important information on the location and relative abundance of proteins inside cells. Gaining access to this information without extracting or disrupting the location of target proteins requires specialized preparation methods. Sectioning frozen blocks of chemically fixed and cryoprotected biological material is one method for obtaining immunocytochemical data. Once the cells or tissues are cut, the thawed cryosections can be labeled with specific antibodies and colloidal gold probes. They are then embedded in a thin film of plastic containing a contrasting agent. Subcellular morphology can be correlated with specific affinity labeling by examination in the transmission electron microscope. Modern technical advancements both in preparation protocols and equipment design make cryosectioning a routine and rapid approach for immunocytochemistry that may provide increased sensitivity for some antibodies.

摘要

用于透射电子显微镜的免疫细胞化学提供了关于细胞内蛋白质位置和相对丰度的重要信息。在不提取或破坏目标蛋白质位置的情况下获取这些信息需要专门的制备方法。切割化学固定和冷冻保护的生物材料的冷冻块是获得免疫细胞化学数据的一种方法。一旦细胞或组织被切割,解冻的冷冻切片可以用特异性抗体和胶体金探针进行标记。然后将它们包埋在含有造影剂的塑料薄膜中。通过在透射电子显微镜下检查,可以将亚细胞形态与特异性亲和标记相关联。制备方案和设备设计方面的现代技术进步使冷冻切片成为免疫细胞化学的常规且快速的方法,这可能会提高某些抗体的灵敏度。

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