Control specimens for immunocytochemistry in liquid‐based cytology

Abstract

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). 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™. 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. 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.