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通过光学显微镜和电子显微镜对副粘病毒抗原进行包埋后免疫细胞化学定位。

Postembedding immunocytochemical localization of paramyxovirus antigens by light and electron microscopy.

作者信息

Schwendemann G, Wolinsky J S, Hatzidimitriou G, Merz D C, Waxham M N

出版信息

J Histochem Cytochem. 1982 Dec;30(12):1313-9. doi: 10.1177/30.12.6296222.

Abstract

A postembedding method is described to localize antigens specific for various paramyxoviruses in sections of cells and tissues that have been fixed and embedded in epoxy resins for conventional electron microscopy. Viral antigens were localized in CV-1 cell cultures infected with simian virus 5 (SV5), brains of suckling hamsters inoculated with either neuroadapted mumps virus or hamster-adapted measles virus, and brains of adult mice infected with Sendai (parainfluenza I) virus. Both 1-micrometer-thick and thin (gold) tissue sections were etched with alcoholic sodium hydroxide-solution and then treated following either the unlabeled antibody peroxidase-antiperoxidase or the biotinylated protein A:avidin peroxidase procedure. Primary reagents included immunoglobulin isolated from hyperimmune rabbit sera with specificity to the major viral components of SV5 or SV5 hemagglutinin-neuraminidase, to whole mumps virus or mumps virus nucleocapsids, and to whole Sendai virus. Crude rabbit anti-Sendai virus antiserum and whole human subacute sclerosing panencephalitis (SSPE) sera were used in parallel. The results indicate that tissues processed for conventional evaluation by electron microscopy may be suitable, within limits, for postembedding immunocytochemical staining of paramyxovirus antigens.

摘要

本文描述了一种包埋后方法,用于在已固定并包埋于环氧树脂中以进行常规电子显微镜检查的细胞和组织切片中定位各种副粘病毒的特异性抗原。病毒抗原定位在感染猿猴病毒5(SV5)的CV-1细胞培养物、接种了神经适应型腮腺炎病毒或仓鼠适应型麻疹病毒的乳鼠脑以及感染仙台(副流感I)病毒的成年小鼠脑中。1微米厚的组织切片和薄(金)组织切片均用乙醇氢氧化钠溶液蚀刻,然后按照未标记抗体过氧化物酶-抗过氧化物酶或生物素化蛋白A:抗生物素蛋白过氧化物酶程序进行处理。主要试剂包括从对SV5主要病毒成分或SV5血凝素-神经氨酸酶、全腮腺炎病毒或腮腺炎病毒核衣壳以及全仙台病毒具有特异性的超免疫兔血清中分离的免疫球蛋白。同时使用粗制兔抗仙台病毒抗血清和全人亚急性硬化性全脑炎(SSPE)血清。结果表明,经常规电子显微镜评估处理的组织在一定限度内可能适用于副粘病毒抗原的包埋后免疫细胞化学染色。

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