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液基细胞学中免疫细胞化学的对照标本。

Control specimens for immunocytochemistry in liquid-based cytology.

作者信息

Hansen T, Pedersen H, Brauner V, Hariri J

机构信息

Department of Pathology, Sygehus Sønderjylland, Sønderborg, Denmark.

出版信息

Cytopathology. 2011 Aug;22(4):243-6. doi: 10.1111/j.1365-2303.2010.00755.x. Epub 2010 May 12.

Abstract

OBJECTIVE

Immunostaining necessitates the use of positive as well as negative controls, which is usually an easy procedure in immunohistochemistry (IHC). To find suitable control specimens for immunocytochemistry (ICC) is, on the other hand, a challenging task and to the best of our knowledge is not sufficiently dealt with in the English literature. The aim of this trial was to develop an applicable method to select, collect, process and store control specimens for ICC using liquid-based cytology (LBC).

METHODS

The study included 21 different antibodies, which were known to react with at least one of the cellular components from tonsils, serous fluids and bronchial washings. The LBC specimens from the tonsils were collected as SurePath™ specimens (BD, Bencton, Dickinson and Company) by brushing the cut-surface of a fresh tonsil and then immersing the brush head into the SurePath™ vial. The serous fluids and bronchial washings were fixed in CytoRich Red™ (BD). Some of the cellular suspensions from the tonsils and equal amounts of the serous fluid and the bronchial washings were also mixed as a cocktail. Unstained SurePath slides were then prepared on the PrepStain™ (BD) Non-GYN Program, and the unstained and dry slides were then stored at 5 °C to test the effect of storage on the preservation of the antigenicity. ICC was then performed on BenchMark-XT™.

RESULTS

Cellular components in unstained SurePath™ slides reacted positively with relevant antibodies. Slides that were stored for up to 40 days did not loose staining intensity.

CONCLUSION

Specimens from body fluids and cell-suspensions that are collected by brushing the cut-surface from different types of fresh tissues or organs can be used as control specimens either separately or as mixtures. Dry and unstained slides can then be prepared and stored in a refrigerator for at least 40 days without loosing staining intensity.

摘要

目的

免疫染色需要使用阳性和阴性对照,这在免疫组织化学(IHC)中通常是一个简单的过程。另一方面,为免疫细胞化学(ICC)寻找合适的对照标本是一项具有挑战性的任务,据我们所知,英文文献中对此并未充分探讨。本试验的目的是开发一种适用的方法,用于使用液基细胞学(LBC)选择、收集、处理和储存ICC的对照标本。

方法

该研究包括21种不同的抗体,已知它们可与扁桃体、浆液和支气管灌洗中的至少一种细胞成分发生反应。通过刷新鲜扁桃体的切面,然后将刷头浸入SurePath™小瓶中,收集扁桃体的LBC标本作为SurePath™标本(BD,本顿迪金森公司)。浆液和支气管灌洗标本用CytoRich Red™(BD)固定。扁桃体的一些细胞悬液以及等量的浆液和支气管灌洗标本也混合成一种混合液。然后在PrepStain™(BD)非妇科程序上制备未染色的SurePath载玻片,将未染色且干燥的载玻片储存在5°C下,以测试储存对抗原性保存的影响。然后在BenchMark-XT™上进行ICC。

结果

未染色的SurePath™载玻片中的细胞成分与相关抗体呈阳性反应。储存长达40天的载玻片没有失去染色强度。

结论

通过刷不同类型新鲜组织或器官的切面收集的体液和细胞悬液标本可单独或混合用作对照标本。然后可以制备干燥且未染色的载玻片,并储存在冰箱中至少40天而不失去染色强度。

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