Cajulis R S, Frias-Hidvegi D, Yu G H, Eggena S
Northwestern University, Chicago, IL 60611-3008, USA.
Diagn Cytopathol. 1996 Mar;14(2):178-81. doi: 10.1002/(SICI)1097-0339(199603)14:2<178::AID-DC14>3.0.CO;2-J.
Fluorescence in situ hybridization (FISH) is rapidly emerging as a tool for analyzing numerical and structural chromosomal abnormalities in both liquid and solid tumors. Most studies make use of fresh samples. To determine the feasibility of detecting numerical chromosomal abnormalities (NCA) by FISH using chromosome-specific probes 8, 12, 17, and X (Vysis, Inc., Downers Grove, IL) on archival cytologic preparations, we studied 23 patient samples, one Papanicolaou-and one Diff-Quik-stained slide per case (46 slides), and two additional unstained slides (fresh ascitic fluids) as controls. Included in this study were nine ascitic fluids (four benign and five malignant), four malignant pleural fluids, three benign bladder washes, and seven malignant fine-needle aspirates (FNA) from various sites. The slides ranged from 1-94 days old. After removal of coverslips using xylene, all slides were destained in a series of alcohol and water washes. Pretreatment of slides with pepsin was followed by the in situ hybridization procedure. Two hundred cells per slide were evaluated for distinct separate signals. Results showed the following: 1) all slides were evaluable except for eight (8/46) which had either too few cells or enough cells but with faint signals, 2) the oldest sample showed distinct signals, 3) previously Diff-Quik-stained slides showed relatively better signals than Papanicolaou-stained slides, 4) samples less than a month old showed relatively better signals, and 5) malignant samples showed various NCA, but not the benign samples. We conclude that FISH on archival cytologic preparation 1) is feasible, although age of the slide is a factor since better signals were seen in those less than a month old, 2) shows better results in previously Diff-Quik-stained slides, and 3) is a tool that can be used in the retrospective study of various liquid and solid neoplasms.
荧光原位杂交（FISH）正迅速成为一种用于分析液体和实体瘤中染色体数目和结构异常的工具。大多数研究使用新鲜样本。为了确定在存档细胞学标本上使用染色体特异性探针8、12、17和X（Vysis公司，伊利诺伊州唐纳斯格罗夫）通过FISH检测染色体数目异常（NCA）的可行性，我们研究了23例患者样本，每例有一张巴氏染色和一张Diff - Quik染色的玻片（共46张玻片），以及另外两张未染色的玻片（新鲜腹水）作为对照。本研究包括9份腹水（4份良性和5份恶性）、4份恶性胸水、3份良性膀胱冲洗液以及7份来自不同部位的恶性细针穿刺抽吸物（FNA）。玻片的保存时间为1至94天。使用二甲苯去除盖玻片后，所有玻片在一系列酒精和水洗中进行脱色。玻片用胃蛋白酶预处理后进行原位杂交程序。每张玻片评估200个细胞的明显独立信号。结果显示如下：1）除8张（8/46）玻片外，所有玻片均可评估，这8张玻片要么细胞数量过少，要么细胞数量足够但信号微弱；2）最陈旧的样本显示出明显的信号；3）先前Diff - Quik染色的玻片显示的信号相对比巴氏染色的玻片更好；4）保存时间不到一个月的样本显示的信号相对更好；5）恶性样本显示出各种NCA，但良性样本未显示。我们得出结论，在存档细胞学标本上进行FISH：1）是可行的，尽管玻片的保存时间是一个因素，因为在保存时间不到一个月的样本中信号更好；2）在先前Diff - Quik染色的玻片中显示出更好的结果；3）是一种可用于各种液体和实体肿瘤回顾性研究的工具。
