Rivasi Francesco, Schirosi Laura, Bettelli Stefania, Bigiani Nazzarena, Curatola Carlo
Department of Diagnostic and Laboratory Services and Forensic Medicine, Section of Pathological Anatomy, University of Modena and Reggio Emilia, Modena, Italy.
Diagn Cytopathol. 2008 Sep;36(9):633-6. doi: 10.1002/dc.20883.
Postmortem studies on still-borns and miscarriages are important to define the sex and eventually the morphologic anomalies correlated to chromosomal aberrations. When the conditions for carrying out a cytogenetic study do not exist, these chromosomal alterations can be investigated by nucleic acid fluorescent in situ hybridization (FISH), which can be performed on interphase nuclei, usually on formalin-fixed paraffin embedded tissues or on fresh cytological specimens. The objective of the present study is to prove whether this technique can be successfully applied to formalin-fixed cell touch preparations and cytological specimens obtained from foetal autopsies. The study was carried out 12 abortions some of which were spontaneous and some of which were therapeutic. The materials were formalin-fixed. Cell touch preparations and cytological specimens were obtained. The FISH was performed using X/Y probes (Vysis) and the Aneuvysion Kit (05J38-030, Vysis), the probes being for chromosomes 13/21 and X/Y/18. To verify the reliability of the technique, the same reactions were also performed on fresh analogous materials. The slides were evaluable, and the probes hybridized to interphase nuclei showed distinct signals. All the samples were adequate for FISH analysis without any notable difference in the results. Moreover, it is technically possible to perform this analysis not only on fresh but particularly on formalin-fixed cytological specimens. On the other hand, the use of this type of cytological samples, as compared to formalin-fixed and paraffin embedded tissue sections, has the advantage of presenting intact, noncut nuclei with preserved cytomorphology, avoiding the problems of overlapping nuclei and making the identification of the real chromosomal arrangement easier.
对死产儿和流产儿进行尸检研究对于确定性别以及最终确定与染色体畸变相关的形态学异常非常重要。当不存在进行细胞遗传学研究的条件时，可以通过核酸荧光原位杂交（FISH）来研究这些染色体改变，该技术可在间期核上进行，通常用于福尔马林固定石蜡包埋组织或新鲜细胞学标本。本研究的目的是证明该技术是否能成功应用于福尔马林固定的细胞印片以及从胎儿尸检获得的细胞学标本。该研究对12例流产病例进行，其中一些是自然流产，一些是治疗性流产。材料均用福尔马林固定。获取了细胞印片和细胞学标本。使用X/Y探针（Vysis）和Aneuvysion试剂盒（05J38 - 030，Vysis）进行FISH，这些探针针对13/21号染色体以及X/Y/18号染色体。为验证该技术的可靠性，还对新鲜的类似材料进行了相同反应。玻片可评估，与间期核杂交的探针显示出清晰的信号。所有样本均适合进行FISH分析，结果无显著差异。此外，不仅在新鲜标本上，而且特别在福尔马林固定的细胞学标本上进行这种分析在技术上是可行的。另一方面，与福尔马林固定石蜡包埋组织切片相比，使用这种类型的细胞学样本具有呈现完整、未切割且细胞形态保存完好的细胞核的优点，避免了细胞核重叠的问题，使确定真实的染色体排列更容易。
