采用荧光原位杂交（Q-FISH）技术对人组织切片中端粒长度的定量测量。

Telomere Length Measurement in Human Tissue Sections by Quantitative Fluorescence In Situ Hybridization (Q-FISH).

机构信息

Research Team for Geriatric Pathology, Tokyo Metropolitan Institute for Geriatrics and Gerontology, Tokyo, Japan.

Department of Pathology, Tokyo Metropolitan Institute for Geriatrics and Gerontology, Tokyo, Japan.

出版信息

Methods Mol Biol. 2025;2857:9-14. doi: 10.1007/978-1-0716-4128-6_2.

DOI:10.1007/978-1-0716-4128-6_2

PMID:39348051

Abstract

Telomeres in most somatic cells shorten with each cell division, and critically short telomeres lead to cellular dysfunction, cell cycle arrest, and senescence. Thus, telomere shortening is an important hallmark of human cellular senescence. Quantitative fluorescence in situ hybridization (Q-FISH) using formalin-fixed paraffin-embedded (FFPE) tissue sections allows the estimation of telomere lengths in individual cells in histological sections. In our Q-FISH method, fluorescently labelled peptide nucleic acid (PNA) probes are hybridized to telomeric and centromeric sequences in FFPE human tissue sections, and relative telomere lengths (telomere signal intensities relative to centromere signal intensities) are measured. This chapter describes our Q-FISH protocols for assessing relative telomere lengths in FFPE human tissue sections.

摘要

端粒在大多数体细胞中随着细胞分裂而缩短，而端粒过短会导致细胞功能障碍、细胞周期停滞和衰老。因此，端粒缩短是人类细胞衰老的一个重要标志。使用福尔马林固定石蜡包埋（FFPE）组织切片的定量荧光原位杂交（Q-FISH）可以估计组织切片中单个细胞的端粒长度。在我们的 Q-FISH 方法中，荧光标记的肽核酸（PNA）探针与 FFPE 人类组织切片中的端粒和着丝粒序列杂交，并测量相对端粒长度（端粒信号强度与着丝粒信号强度的比值）。本章介绍了我们用于评估 FFPE 人类组织切片中相对端粒长度的 Q-FISH 方案。