Abstract：
Single-molecule fluorescence in situ hybridization (smFISH) represents a promising approach for the quantitative analysis of nucleic acid biomarkers in clinical tissue samples. However, low signal intensity and high background noise are complications that arise from diagnostic pathology when performed with smFISH-based RNA imaging in formalin-fixed paraffin-embedded (FFPE) tissue specimens. Moreover, the associated complex procedures can produce uncertain results and poor image quality. Herein, by combining the high specificity of split DNA probes with the high signal readout of ZnCdSe/ZnS quantum dot (QD) labeling, we introduce QD split-FISH, a high-brightness smFISH technology, to quantify the expression of mRNA in both cell lines and clinical FFPE tissue samples of breast cancer and lung squamous carcinoma. Owing to its high signal-to-noise ratio, QD split-FISH is a fast, inexpensive, and sensitive method for quantifying mRNA expression in FFPE tumor tissues, making it suitable for biomarker imaging and diagnostic pathology.
摘要：
单分子荧光原位杂交（smFISH）代表了用于临床组织样品中核酸生物标志物定量分析的有前途的方法。然而,在福尔马林固定石蜡包埋(FFPE)组织标本中使用基于smFISH的RNA成像时,低信号强度和高背景噪声是诊断病理学引起的并发症.此外，相关的复杂程序可能会产生不确定的结果和较差的图像质量。在这里，通过将分裂DNA探针的高特异性与ZnCdSe/ZnS量子点（QD）标记的高信号读出相结合，我们介绍QDsplit-FISH，高亮度smFISH技术，定量乳腺癌和肺鳞癌细胞系和临床FFPE组织样本中mRNA的表达。由于其高信噪比，QD分裂-FISH是一个快速，便宜,和定量FFPE肿瘤组织中mRNA表达的灵敏方法，使其适用于生物标志物成像和诊断病理学。
