幻灯片上哺乳动物细胞的多重定量原位杂交：qHCR 和 dHCR 成像（v3.0）。

Multiplexed Quantitative In Situ Hybridization for Mammalian Cells on a Slide: qHCR and dHCR Imaging (v3.0).

机构信息

Division of Biology & Biological Engineering, California Institute of Technology, Pasadena, CA, USA.

Division of Engineering & Applied Science, California Institute of Technology, Pasadena, CA, USA.

出版信息

Methods Mol Biol. 2020;2148:143-156. doi: 10.1007/978-1-0716-0623-0_9.

DOI:10.1007/978-1-0716-0623-0_9

PMID:32394380

Abstract

In situ hybridization based on the mechanism of hybridization chain reaction (HCR) enables multiplexed quantitative mRNA imaging in diverse sample types. Third-generation in situ HCR (v3.0) provides automatic background suppression throughout the protocol, dramatically enhancing performance and ease of use. In situ HCR v3.0 supports two quantitative imaging modes: (1) qHCR imaging for analog mRNA relative quantitation with subcellular resolution and (2) dHCR imaging for digital mRNA absolute quantitation with single-molecule resolution. Here, we provide protocols for qHCR and dHCR imaging in mammalian cells on a slide.

摘要

基于杂交链式反应（HCR）机制的原位杂交使在多种样本类型中进行多重定量 mRNA 成像成为可能。第三代原位 HCR（v3.0）在整个方案中提供自动背景抑制，极大地提高了性能和易用性。原位 HCR v3.0 支持两种定量成像模式：（1）qHCR 成像，用于具有亚细胞分辨率的模拟 mRNA 相对定量，（2）dHCR 成像，用于具有单分子分辨率的数字 mRNA 绝对定量。本文提供了在载玻片上进行哺乳动物细胞 qHCR 和 dHCR 成像的方案。