Dinges H P, Schmid C, Zatloukal K, Mair S, Preisegger K H, Redl H
Institute of Pathology, University of Graz School of Medicine, Austria.
Pathol Res Pract. 1992 Aug;188(6):714-21. doi: 10.1016/S0344-0338(11)80167-6.
A method has been established for storage and preservation of cytological specimens in liquid nitrogen and further processing for immunocytochemistry as smears prepared from thawed cells or cryo-sections of frozen cell pellets. For the experiments cultured cells of a T-lymphoblastic leukemia cell line (ATCC CCL 119) and blood cells of the buffy coat of healthy humans were treated with a cryo-solution (fetal calf serum +5% dimethylsulfoxid) and after freezing stored in liquid nitrogen. Alternatively, cells preincubated with cryo-solution followed by suspension in fetal calf serum without cryo-additive were frozen and stored in liquid nitrogen for the production of cryo-sections. Indirect immunofluorescence and alkaline phosphatase--antialkaline phosphatase based immunoreactions were performed for the decoration of various surface antigens with a panel of monoclonal antibodies. All immunoreactions were repeated at least three times and the stored cell preparations were investigated after different periods of storage (up to four months). The immunoreactions of fresh cells in suspension (which were used as controls) were comparable with those of cryopreserved cells, e.g. cells on smears after thawing and on cryo-sections of cell pellets. The strongest immunoreactions were achieved on fixed cryo-sections. The maintenance of cell morphology of smears from cryopreserved cells was slightly better than of cells from cryo-sections. In our hands the preparation of cell pellets, which are suitable for the storage in liquid nitrogen and the production of cryosections, is a very useful method for immunocytochemical investigations of cytological specimens especially in situations where immunoreactions cannot be performed on fresh material.(ABSTRACT TRUNCATED AT 250 WORDS)
已建立一种方法，用于将细胞学标本储存在液氮中，并进一步处理以进行免疫细胞化学分析，即从解冻细胞制备涂片或对冷冻细胞沉淀进行冰冻切片。对于实验，将T淋巴细胞白血病细胞系（ATCC CCL 119）的培养细胞和健康人血沉棕黄层的血细胞用冷冻溶液（胎牛血清+5%二甲基亚砜）处理，冷冻后储存在液氮中。或者，将预先用冷冻溶液孵育、然后悬浮于无冷冻添加剂的胎牛血清中的细胞冷冻并储存在液氮中，用于制备冰冻切片。使用一组单克隆抗体通过间接免疫荧光和基于碱性磷酸酶-抗碱性磷酸酶的免疫反应来检测各种表面抗原。所有免疫反应至少重复三次，并在不同储存期（长达四个月）后对储存的细胞制剂进行研究。悬浮新鲜细胞（用作对照）的免疫反应与冷冻保存细胞的免疫反应相当，例如解冻后涂片上的细胞和细胞沉淀的冰冻切片上的细胞。在固定的冰冻切片上获得最强的免疫反应。冷冻保存细胞涂片的细胞形态维持略优于冰冻切片的细胞。在我们看来，制备适合储存在液氮中并用于制作冰冻切片的细胞沉淀，对于细胞学标本的免疫细胞化学研究是一种非常有用的方法，尤其是在无法对新鲜材料进行免疫反应的情况下。（摘要截短于250字）
