开发一种样本制备方法,用于对甲固定型胺嵌入型肝组织样本进行微蛋白质学分析
Yong-Er Wang1, Wei-Lan Zeng1, Sheng-Tian Cao1
1Biomedical Research Center, Southern Medical University, Guangzhou, China; School of Pharmaceutical Science, Southern Medical University, Guangzhou, China.
Talanta
|August 28, 2023
概括
这项研究引入了一种新方法,用于从甲固定嵌 (FFPE) 组织中准备微小的肝脏样本,用于质谱 (MS) 分析. 开发的协议HDMSP提高了FFPE肝脏微蛋白质的蛋白质提取效率和精度.
科学领域:
- 生物化学 生物化学
- 蛋白质组学是指蛋白质组学.
- 分子生物学分子生物学
背景情况:
- 嵌入式甲固化 (FFPE) 的样本对肝脏研究有价值,但对微型蛋白质构成挑战.
- 现有的准备FFPE样本用于质谱 (MS) 的方法通常是低效和耗时的.
研究的目的:
- 使用FFPE肝脏样本开发微型蛋白质学优化样品制备方法.
- 为了提高蛋白质提取,恢复和MS分析精度,对有限的FFPE肝脏样本进行.
主要方法:
- 开发了一种新的协议,HDMSP (高效消化和基于磁珠的样品制备),用于从FFPE肝脏样本中微量提取蛋白质.
- 利用激光捕获微解剖来从FFPE肝脏切片中每批收集2000个单个细胞.
- 优化蛋白质提取使用性氨基缓冲剂和二硫酸盐,然后以磁珠为基础的沉用于MS分析.
主要成果:
- 在HDMSP协议中,与标准方法相比,FFPE样本的蛋白质回收率 (PRR) 显著提高,为88.8%.
- 使用HDMSP进行的MSS分析显示,FFPE肝脏微蛋白质学的深度,稳定性和定量准确性增加.
- 经过HDMSP处理的FFPE样本表现出与使用过器辅助样本制备 (FASP) 处理的新鲜冷样本相似的物理化学特性和亚细胞位置.
- 与FASP相比,HDMSP减少了15.9%的样本准备时间和30.8%的实验成本.
结论:
- HDMSP是一种简单,有效和具有成本效益的FFPE肝脏微蛋白学方法.
- 这种方法可以从有限的FFPE肝脏样本中进行高质量的蛋白质组分析,从而推动肝脏研究.
- HDMSP为研究人员提供了一种有价值的解决方案,用于使用FFPE组织进行基于质谱的蛋白质组学研究.
更多相关视频
08:16Optimal Preparation of Formalin Fixed Samples for Peptide Based Matrix Assisted Laser Desorption/Ionization Mass Spectrometry Imaging Workflows
Published on: January 16, 2018
9.0K
09:00A Streamlined Approach for Mass Spectrometry-Based Proteomics Using Selected Tissue Regions
Published on: April 18, 2025
702
相关概念视频
01:20Preparation of Samples for Electron Microscopy
5.5K
To be visualized by an electron microscope, either transmission or scanning, biological samples need to be fixed (stabilized) so the electron beam does not destroy them and dried thoroughly (desiccated/dehydrated) so the vacuum does not affect them. Fixation needs to be done as quickly as possible because the sample properties will start changing as soon as it is removed from its natural environment. For example, in a tissue sample, the oxygen levels begin decreasing, causing an altered...
5.5K
01:03Fixation and Sectioning
4.4K
Two basic types of preparation are used to visualize specimens with a light microscope: wet mounts and fixed specimens.
The simplest type of preparation is the wet mount, in which the specimen is placed in a drop of liquid on the slide. A liquid specimen can be directly deposited on the slide using a dropper. Solid specimens, such as skin scraping, can be placed on the slide before adding a drop of liquid to prepare the wet mount. Sometimes the liquid is simply water, but stains are often added...
The simplest type of preparation is the wet mount, in which the specimen is placed in a drop of liquid on the slide. A liquid specimen can be directly deposited on the slide using a dropper. Solid specimens, such as skin scraping, can be placed on the slide before adding a drop of liquid to prepare the wet mount. Sometimes the liquid is simply water, but stains are often added...
4.4K