利用 pSABER 对细胞和组织病理学样本中的核酸靶标进行高效和高度扩增成像。

IF 36.1 1区 生物学 Q1 BIOCHEMICAL RESEARCH METHODS
Sahar Attar, Valentino E Browning, Mary Krebs, Yuzhen Liu, Eva K Nichols, Ashley F Tsue, David M Shechner, Jay Shendure, Joshua A Lieberman, Devin K Schweppe, Shreeram Akilesh, Brian J Beliveau
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引用次数: 0

摘要

原位杂交(ISH)是研究固定样本中核酸靶标空间排列的有力工具。ISH 通常使用荧光团进行可视化,以实现高灵敏度和多路复用,或使用比色标签,以方便与组织病理学染色共可见。这两种方法都得益于信号放大,从而使目标检测有效、快速,并与多种光学系统兼容。pSABER 将辣根过氧化物酶共轭寡核苷酸的结合位点装饰在原位靶点上,从而实现荧光或比色基质的局部沉积。我们证明 pSABER 能有效标记培养细胞和 FFPE 标本中的 DNA 和 RNA 靶标。此外,与传统的交换反应信号放大(SABER)相比,pSABER 可实现五倍的信号放大,并可通过溶液交换进行串行复用。因此,pSABER 将核酸检测与能够提供多种读数的强大信号放大技术相结合,将在研究和临床中发挥广泛的作用。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
Efficient and highly amplified imaging of nucleic acid targets in cellular and histopathological samples with pSABER.

In situ hybridization (ISH) is a powerful tool for investigating the spatial arrangement of nucleic acid targets in fixed samples. ISH is typically visualized using fluorophores to allow high sensitivity and multiplexing or with colorimetric labels to facilitate covisualization with histopathological stains. Both approaches benefit from signal amplification, which makes target detection effective, rapid and compatible with a broad range of optical systems. Here, we introduce a unified technical platform, termed 'pSABER', for the amplification of ISH signals in cell and tissue systems. pSABER decorates the in situ target with concatemeric binding sites for a horseradish peroxidase-conjugated oligonucleotide, enabling the localized deposition of fluorescent or colorimetric substrates. We demonstrate that pSABER effectively labels DNA and RNA targets in cultured cells and FFPE specimens. Furthermore, pSABER can achieve fivefold signal amplification over conventional signal amplification by exchange reaction (SABER) and can be serially multiplexed using solution exchange. Therefore, by linking nucleic acid detection to robust signal amplification capable of diverse readouts, pSABER will have broad utility in research and clinical settings.

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来源期刊
Nature Methods
Nature Methods 生物-生化研究方法
CiteScore
58.70
自引率
1.70%
发文量
326
审稿时长
1 months
期刊介绍: Nature Methods is a monthly journal that focuses on publishing innovative methods and substantial enhancements to fundamental life sciences research techniques. Geared towards a diverse, interdisciplinary readership of researchers in academia and industry engaged in laboratory work, the journal offers new tools for research and emphasizes the immediate practical significance of the featured work. It publishes primary research papers and reviews recent technical and methodological advancements, with a particular interest in primary methods papers relevant to the biological and biomedical sciences. This includes methods rooted in chemistry with practical applications for studying biological problems.
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