Objective: Although transcriptomic assessments of small samples using high-throughput techniques are usually performed on fresh or frozen tissues, there is a growing demand for those performed on stained cellular specimens already used for diagnostic purposes. Study Design: The possibility of detecting mRNAs and microRNAs (miRNAs) from routinely processed cytological samples using nCounter® technology was explored. Fresh samples from pleural and peritoneal effusions were analyzed using 2 parallel methods: samples were smeared and routinely stained using the May-Grünwald-Giemsa or Diff-Quik® method and mounted using conventional methods, and they were also studied following a snap freezing method, in which samples were maintained at −80°C until use. mRNAs and miRNAs were assessed and compared after total RNA extraction from both routinely processed samples and their matched frozen controls. Results: A good concordance was found between the gene expression measured in routinely processed samples and their matched frozen controls for the majority of mRNAs and miRNAs tested. However, the standard deviation of low-expressed miRNA was high. Conclusions: Although nCounter® technology is a robust method to measure and characterize both mRNAs and miRNAs from routinely processed cytological samples, caution is recommended for the interpretation of low-expressed miRNA.
Acta Cytologica

中文翻译：

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常规处理和染色细胞学涂片中mRNA和miRNA的数字多路复用基因表达分析：原理验证研究

目的：尽管通常在新鲜或冷冻的组织上使用高通量技术对小样本进行转录组评估，但对已经用于诊断目的的染色细胞样本进行转录组评估的需求不断增长。学习规划：探索了使用nCounter®技术从常规处理的细胞学样品中检测mRNA和microRNA（miRNA）的可能性。使用两种平行方法分析了来自胸膜和腹膜积液的新鲜样品：使用May-Grünwald-Giemsa或Diff-Quik®方法对样品进行涂片和常规染色，并使用常规方法进行固定，还采用快速冷冻方法进行了研究，其中样品保持在-80°C直至使用。从常规处理的样品及其匹配的冷冻对照中提取总RNA后，评估并比较mRNA和miRNA。结果：在常规处理的样品中测得的基因表达与测试的大多数mRNA和miRNA的匹配冷冻对照之间发现了很好的一致性。但是，低表达的miRNA的标准差较高。结论：尽管nCounter®技术是一种测量和表征常规处理的细胞学样品中的mRNA和miRNA的可靠方法，但建议对低表达miRNA的解释要谨慎。
细胞学学报