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相关实验视频

Updated: May 2, 2026

Quantitative Analysis of Alternative Pre-mRNA Splicing in Mouse Brain Sections Using RNA In Situ Hybridization Assay
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来自小鼠的甲固定嵌肌肉骨组织样本的准备协议,用于空间转录学.

Esther Wehrle1, Denise Günther2, Neashan Mathavan2

  • 1Institute for Biomechanics, ETH Zurich, Zurich, Switzerland; AO Research Institute Davos, Davos Platz, Switzerland.

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|March 31, 2024
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概括

这项研究详细介绍了在小鼠骨和肌肉骨组织上进行空间转录的新协议. 该方法可以对甲固定嵌入的样品进行详细的分子分析.

关键词:
基因组学就是基因组学.健康科学 卫生科学 卫生科学显微镜的使用方法分子生物学分子生物学分子/化学探头 分子/化学探头

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科学领域:

  • 分子生物学分子生物学
  • 基因组学就是基因组学.
  • 生物技术是生物技术.

背景情况:

  • 空间转录学为组织结构和细胞功能提供了前所未有的洞察力.
  • 分析骨和肌肉骨组织由于其复杂的组成和矿物化而带来了独特的挑战.
  • 嵌入式甲固定 (FFPE) 样本对于回顾性研究是有价值的,但需要用于转录基因分析的特殊处理.

研究的目的:

  • 建立并提出一个全面的协议,用于应用空间转录学到小鼠骨和多组织肌肉骨FFPE样本.
  • 提供详细的程序步骤,从组织采集到数据分析.
  • 解决与FFPE肌肉骨组织相关的特定技术障碍.

主要方法:

  • 在小鼠肌肉骨样本中采集组织,加工和对嵌.
  • 切割FFPE块并放置在专门的幻灯片上进行空间转录.
  • 解链,RNA库的准备,以及随后的高通量测序.
  • 生物信息分析管道用于空间转录基因组数据的解释.

主要成果:

  • 一个完整的18天的协议是描述的小鼠大腿骨和相邻的肌肉FFPE样本.
  • 超过50%的协议时间用于矿化骨组织的脱.
  • 该协议从具有挑战性的FFPE肌肉骨样本中获得高质量的空间转录基因数据.

结论:

  • 该协议促进了小鼠骨和肌肉骨组织的高分辨率空间转录组分析.
  • 该方法对于理解复杂解剖结构中的组织异质性和细胞相互作用至关重要.
  • 它扩大了存档的FFPE样本的实用性,用于肌肉骨生物学中的先进分子研究.