使用MALDI质谱成像和现场光标记以高空间分辨率可视化代谢物和微生物
Patric Bourceau1,2, Benedikt Geier1,3, Vincent Suerdieck4
1Max Planck Institute for Marine Microbiology, Bremen, Germany.
Nature protocols
|September 6, 2023
概括
我们开发了metaFISH,这是一种结合质谱成像和光 in situ 杂交的方法,用于绘制组织中的代谢物和微生物的地图. 这种方法允许单细胞解决宿主-微生物代谢相互作用.
科学领域:
- 微生物学 微生物学
- 生物化学 生化学
- 分子成像学分子成像学
背景情况:
- 无标签的分子成像,例如矩阵辅助激光脱离离电离质谱成像 (MALDI-MSI),可以同时绘制组织中众多代谢物的图像.
- 在宿主-微生物相互作用中定位微生物和将代谢物分配给宿主或微生物组成员仍然具有挑战性.
研究的目的:
- 开发一种关联成像方法,将MALDI-MSI与光 in situ杂交 (FISH) 结合起来,用于精确的微生物定位和代谢物分配.
- 引入metaFISH作为与微生物组成员相关的空间代谢物分布的单细胞分辨的可靠方法.
主要方法:
- 用核酸探针进行组织制备和组织上的杂交.
- 光显微镜用于微生物识别和定位.
- MALDI-MSI数据采集和与FISH成像进行整合,以进行相关分析.
主要成果:
- metaFISH可以在单个组织段上以微米分辨率绘制数百种代谢物和多种微生物的地图.
- 该方法成功地将细胞内和细胞外细菌,宿主细胞及其相关代谢物定位在动物组织中.
- 低丰度细菌感染部位的识别有助于高分辨率MSI分析.
结论:
- metaFISH提供了一种强大的工具,以单细胞分辨率剖析宿主及其微生物群之间的复杂代谢相互作用.
- 该协议是用户友好的,需要大约两个工作日,适用于从环境微生物学到临床科学的各种研究领域.
更多相关视频
09:49Investigation of Microbial Cooperation via Imaging Mass Spectrometry Analysis of Bacterial Colonies Grown on Agar and in Tissue During Infection
Published on: November 18, 2022
2.1K
09:31Visualization of Gut Microbiota-host Interactions via Fluorescence In Situ Hybridization, Lectin Staining, and Imaging
Published on: July 9, 2021
8.3K
相关概念视频
01:37Super-resolution Fluorescence Microscopy
7.0K
Super-resolution fluorescence microscopy (SRFM) provides a better resolution than conventional fluorescence microscopy by reducing the point spread function (PSF). PSF is the light intensity distribution from a point that causes it to appear blurred. Due to PSF, each fluorescing point appears bigger than its actual size, and it is the PSF interference of nearby fluorophores that causes the blurred image. Various approaches to achieving higher resolution through SRFM have recently been...
7.0K
01:28Three-Dimensional Microscopy in Microbiology
65
Three-dimensional imaging techniques are essential in cell biology, allowing researchers to visualize intricate cellular structures with high resolution. Two prominent methods, Differential Interference Contrast Microscopy (DIC) and Confocal Scanning Laser Microscopy (CSLM), provide distinct advantages for imaging live and thick specimens, respectively.Differential Interference Contrast MicroscopyDIC microscopy enhances contrast in transparent, unstained samples by converting phase...
65