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[应用免疫组织化学方法对微波固定的研究]

[Study of the microwave fixation using the immunohistochemical method].

作者信息

Hemmi A, Yamaguchi H, Seyama Y, Mori Y

机构信息

Department of Pathology, Koshigaya Hospital, Dokkyo University School of Medicine.

出版信息

Rinsho Byori. 1990 Feb;38(2):193-200.

PMID:1691799
Abstract

We evaluated microwave fixation of formalin immersed and microwave irradiated kidney and liver tissues by keratin immunostain using the paraffin embedded section. In the microwave irradiated tissues, the formalin fixed areas and the alcohol fixed areas were clearly detected because; 1) Alcohol fixed tissues are easily digested by pepsinization but formalin fixed tissues are not, and 2) Formalin fixed tissues revealed intense keratin staining after pepsinization, whereas intense keratin staining were noted in alcohol fixed tissues without preliminary pepsinization. The microwave irradiation fixed completely the tissues in about 20 sec. A thin, formalin-fixed layer was observed in the periphery of the tissues. A thin, alcohol-fixed layer was observed beneath the peripheral formalin fixed layer in some specimens. This area was thought to have been fixed during the dehydration of the paraffin blocks. However with microwave fixation, most of the central areas differed from those of the formalin or alcohol fixed areas. They showed sharp contrast on hematoxylin-eosin staining, conspicuous cellular membrane and only slight cellular shrinkage and swelling. Unlike the formalin and alcohol fixed tissues, the intensity of keratin staining was independent of preliminary pepsinization and keratin staining was observed. Therefore, the central areas were thought to be fixed by the microwave irradiation. This fixation method seems to be useful for immunostaining.

摘要

我们通过使用石蜡包埋切片进行角蛋白免疫染色,评估了经福尔马林浸泡和微波照射的肾脏和肝脏组织的微波固定效果。在微波照射的组织中,福尔马林固定区域和酒精固定区域清晰可辨,原因如下:1)酒精固定的组织经胃蛋白酶消化后易于分解,而福尔马林固定的组织则不然;2)福尔马林固定的组织经胃蛋白酶消化后显示出强烈的角蛋白染色,而未经预先胃蛋白酶消化的酒精固定组织中也观察到强烈的角蛋白染色。微波照射在约20秒内可完全固定组织。在组织周边观察到一层薄的福尔马林固定层。在一些标本中,在周边福尔马林固定层下方观察到一层薄的酒精固定层。该区域被认为是在石蜡块脱水过程中固定的。然而,通过微波固定,大多数中央区域与福尔马林或酒精固定区域不同。它们在苏木精-伊红染色上显示出明显对比,细胞膜明显,细胞仅轻微收缩和肿胀。与福尔马林和酒精固定组织不同,角蛋白染色强度与预先胃蛋白酶消化无关,且观察到角蛋白染色。因此,中央区域被认为是通过微波照射固定的。这种固定方法似乎对免疫染色有用。

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