Detection of plasma cell immunoglobulins in tissue sections optimally fixed for ultrastructural immunocytochemistry.
Detection of plasma cell immunoglobulins in tissue sections optimally fixed for ultrastructural immunocytochemistry.
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DOI：
10.1177/35.2.3098833
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发表时间：
1987-02
期刊：
影响因子：
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通讯作者：
N. Gonatas;J. Gonatas;A. Stieber;T. Ternynck;S. Avrameas
中科院分区：
文献类型：
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作者：
N. Gonatas;J. Gonatas;A. Stieber;T. Ternynck;S. Avrameas
我们描述了浆细胞免疫球蛋白在腘淋巴结振动切片的超微结构定位。用戊二醛-多聚甲醛固定比多聚甲醛或高碘酸赖氨酸-多聚甲醛更好地保存组织和抗原;羊抗小鼠IgG-链霉亲和素-生物素化辣根过氧化物酶（HRP）或Fab-HRP缀合物的生物素化Fab片段得到类似的结果。使用这两种免疫试剂，在用振动切片机切片的第一层细胞中观察到良好的组织保存和抗原检测。抗小鼠IgG与HRP的缀合物未显示任何染色。过氧化物酶染色可见于核膜、粗面内质网池和高尔基体复合体。在高尔基体，染色一致地看到在cis面和相邻的囊泡的池; transcisterbrain显示弱或没有染色，和相邻的囊泡，“包被”囊泡，和颗粒没有染色。这项研究表明，在用戊二醛-多聚甲醛固定的组织中，通过光和超微结构免疫细胞化学进行高质量的组织保存和抗原检测，然后进行振动切片和Fab-生物素-链霉抗生物素蛋白-生物素-HRP或Fab-HRP免疫染色是可行的。
We describe the ultrastructural localization of plasma cell immunoglobulins in vibratome sections of popliteal lymph nodes. Fixation with glutaraldehyde-paraformaldehyde gave better tissue and antigen preservation than paraformaldehyde or periodic acid lysine-paraformaldehyde; biotinylated Fab fragments of sheep anti-mouse IgG-streptavidin-biotinylated horseradish peroxidase (HRP) or Fab-HRP conjugates gave similar results. With both immunoreagents, excellent tissue preservation and antigen detection was observed in the first layer of cells sectioned with the vibratome. Conjugates of anti-mouse IgG with HRP did not show any staining. Peroxidase stain was observed in the nuclear envelope, cisternae of the rough endoplasmic reticulum, and the Golgi apparatus complex. In the Golgi apparatus, staining was seen consistently in cisternae of the cis face and in adjacent vesicles; the trans cisternae showed weak or no stain, and adjacent vesicles, "coated" vesicles, and granules were not stained. This study shows that high quality of tissue preservation and antigen detection, by both light and ultrastructural immunocytochemistry, is feasible in tissue fixed with glutaraldehyde-paraformaldehyde followed by vibratome sectioning and immunostaining with Fab-biotin-streptavidin-biotin-HRP, or Fab-HRP.