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相关概念视频

FISH - Fluorescent In-situ Hybridization02:07

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,...
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相关实验视频

Updated: Jan 16, 2026

Workflow for High-content, Individual Cell Quantification of Fluorescent Markers from Universal Microscope Data, Supported by Open Source Software
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使用开源工具对多重光装置杂交数据的量化和分析.

Kaitlin E Sullivan1, Margarita Kapustina1, Brianna N Bristow1

  • 1Department of Cellular and Physiological Science, University of British Columbia, Vancouver, BC V6T 1Z3, Canada.

STAR protocols
|October 5, 2025
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概括

本研究介绍了FijiFISH和RUHi,它们是开源工具,用于分析多重光在位杂交 (mFISH) 数据. 这些工具能够进行详细的空间基因表达量化和多式单细胞洞察.

关键词:
生物信息学是一种生物信息学.在现场混合化.显微镜的使用方法

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科学领域:

  • 分子生物学分子生物学
  • 生物信息学是一种生物信息学.
  • 图像分析 图像分析

背景情况:

  • 多复合光 in situ 杂交 (mFISH) 是一种强大的技术,用于可视化组织内的基因表达模式.
  • 分析复杂的mFISH数据需要强大的计算工具来准确量化和解释.
  • 现有的方法可能缺乏用于全面的空间基因表达分析所需的灵活性或可访问性.

研究的目的:

  • 为量化和分析多重光 in situ 杂交 (mFISH) 数据提出一个标准化的协议.
  • 介绍和展示两个开源软件工具FijiFISH和RUHi的实用性,用于mFISH数据分析.
  • 为了使多式,单细胞分辨率洞察空间基因表达,包括实验诱导的外源光体.

主要方法:

  • 使用基于ImageJ的插件FijiFISH进行基本的图像处理步骤,包括注册,细胞细分和基因表达量化.
  • 采用RUHi,一个基于R的包,用于高级数据分析,包括通过代码和Shiny应用程序减少维度,集群和可视化.
  • 集成实验数据与外源光体,以实现多模式空间基因表达分析.

主要成果:

  • 通过FijiFISH和RUHi管道成功量化和分析了mFISH数据.
  • 通过FijiFISH实现了精确的细胞细分和基因表达量化.
  • 使用RUHi证明了空间基因表达数据的有效维度减少,聚类和可视化.
  • 提供了多模式,单细胞分辨率洞察空间基因表达模式.

结论:

  • 斐济FISH和RUHi协议为mFISH数据分析提供了一个全面和可访问的解决方案.
  • 这种方法有助于更深入地了解单细胞层面的空间基因表达.
  • 这些工具的开源性质促进了空间转录学领域的更广泛采用和进一步发展.