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Infrared Laser Ablation Sample Transfer for MALDI Imaging

机译:MALDI成像的红外激光烧蚀样品转移

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摘要

An infrared laser was used to ablate material from tissue sections under ambient conditions for direct collection on a matrix assisted laser desorption ionization (MALDI) target. A 10 (mu)m thick tissue sample was placed on a microscope slide and was mounted tissue-side down between 70 and 450 (mu)m from a second microscope slide. The two slides were mounted on a translation stage, and the tissue was scanned in two dimensions under a focused mid-infrared (IR) laser beam to transfer material to the target slide via ablation. After the material was transferred to the target slide, it was analyzed using MALDI imaging using a tandem time-of-flight mass spectrometer. Images were obtained from peptide standards for initial optimization of the system and from mouse brain tissue sections using deposition either onto a matrix precoated target or with matrix addition after sample transfer and compared with those from standard MALDI mass spectrometry imaging. The spatial resolution of the transferred material is approximately 400 (mu)m. Laser ablation sample transfer provides several new capabilities not possible with conventional MALDI imaging including (1) ambient sampling for MALDI imaging, (2) area to spot concentration of ablated material, (3) collection of material for multiple imaging analyses, and (4) direct collection onto nanostructure assisted laser desorption ionization (NALDI) targets without blotting or ultrathin sections.
机译:在环境条件下,使用红外激光烧蚀组织切片中的材料,以直接收集在基质辅助激光解吸电离(MALDI)靶标上。将10μm厚的组织样品放置在显微镜载玻片上,并从第二个显微镜载玻片上将组织面朝下安装在70至450μm之间。将两个载玻片安装在平移台上,并在聚焦的中红外(IR)激光束下二维扫描组织,以通过消融将材料转移至目标载玻片。将材料转移到目标载玻片上后,使用串联飞行时间质谱仪进行MALDI成像分析。图像是从用于系统初始优化的肽标准品和小鼠脑组织切片中获得的,这些样品使用沉积在基质预涂覆的靶标上或在样品转移后加入基质并与标准MALDI质谱成像相比进行了比较。转移的材料的空间分辨率约为400μm。激光烧蚀样品转移提供了传统MALDI成像无法实现的几种新功能,包括(1)用于MALDI成像的环境采样,(2)烧蚀材料的面积至斑点浓度,(3)用于多次成像分析的材料收集以及(4)直接收集到纳米结构辅助的激光解吸电离(NALDI)目标上,而不会产生印迹或超薄切片。

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