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在固定组织培养细胞和组织中对染色体扩散进行SABER-FISH

Sahar Attar¹, Mary Krebs¹, Eva K Nichols¹

¹Department of Genome Sciences, University of Washington, Seattle, WA, USA.

Methods in molecular biology (Clifton, N.J.)

|August 30, 2025

概括

通过交换反应信号放大 (SABER) 显著增强光现场杂交 (FISH) 信号,从而能够检测较小的核酸标. 这种进步克服了传统的FISH限制,改善了RNA和DNA成像.

科学领域:

分子生物学
遗传学
生物技术

背景情况:

在固定样本中可视化核酸序列的过程中,现场光混合化 (FISH) 是至关重要的.
传统的 FISH 信号很低,只能在很大区域或高副本数序列中检测目标.
需要信号放大方法来提高FISH的灵敏度,并扩大目标区域.

研究的目的:

通过交换反应 (SABER) 详细说明信号放大 FISH 技术.
为了证明SABER对增强RNA和DNA检测的能力.
为各种生物样本提供SABER FISH的实用方案.

主要方法:

SABER FISH使用长时间重复的FISH探针.
这些合体招募多个互补的光探测器, 放大信号.
该方法应用于人体和小鼠标本的基相扩散,细胞和各种组织类型 (FFPE,冷切割).

主要成果:

SABER FISH可以提高RNA和DNA信号的5倍至450倍.
这种放大可以准以前无法检测到的较小的核酸区域.
该技术被证明是多重成像的快速,多功能和成本效益.

结论:

与传统的FISH相比,SABER FISH显著提高了核酸检测的灵敏度.
这种方法克服了传统FISH的局限性,使得采样效率高.
SABER FISH为研究不同样本中的核酸定位和丰富性提供了强大的工具.

关键词:

染色体鱼类有氧核酸探针信号放大空间基因组学空间转录学

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FISH - Fluorescent In-situ Hybridization

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Fluorescence in situ hybridization, or FISH, was developed in the early 1980s and has quickly become one of the most widely used techniques in cytogenetics. Labeled probes are used to bind complementary DNA or RNA sequences on a chromosome or in a region within a cell. Earlier, the probes could only be obtained by cloning or reverse transcription of a DNA template. Currently, the probe oligonucleotides can be synthesized synthetically. Additionally, with the advancement of optical techniques,...