Department of Chemical Physics, Faculty of Science and Technology, University of the Basque Country, Bº Sarriena s/n, 48940 Leioa, Spain.
J Am Soc Mass Spectrom. 2011 Feb;22(2):329-38. doi: 10.1007/s13361-010-0024-5. Epub 2011 Jan 28.
Molecular mass images of tissues will be biased if differences in the physicochemical properties of the microenvironment affect the intensity of the spectra. To address this issue, we have performed-by means of MALDI-TOF mass spectrometry-imaging on slices and lipidomic analysis in extracts of frontal cortex, both from the same postmortem tissue samples of human brain. An external calibration was used to achieve a mass accuracy of 10 ppm (1σ) in the spectra of the extracts, although the final assignment was based on a comparison with previously reported species. The spectra recorded directly from tissue slices (imaging) show excellent s/n ratios, almost comparable to those obtained from the extracts. In addition, they retain the information about the anatomical distribution of the molecular species present in autopsied frozen tissue. Further comparison between the spectra from lipid extracts devoid of proteins and those recorded directly from the tissue unambiguously show that the differences in lipid composition between gray and white matter observed in the mass images are not an artifact due to microenvironmental influences of each anatomical area on the signal intensity, but real variations in the lipid composition.
如果微环境的物理化学性质差异影响光谱强度,那么组织的分子质量图像将存在偏差。为了解决这个问题,我们通过 MALDI-TOF 质谱成像技术对切片进行了研究,并对提取的额皮质进行了脂质组学分析,这些样本均来自于相同的人脑死后组织。采用外部校准来实现提取光谱中 10ppm(1σ)的质量精度,尽管最终的分配是基于与先前报道的物质的比较。直接从组织切片(成像)记录的光谱显示出极好的信噪比,几乎可与从提取物中获得的信噪比相媲美。此外,它们保留了存在于尸检冷冻组织中分子物种的解剖分布信息。对不含蛋白质的脂质提取物的光谱与直接从组织记录的光谱进行进一步比较,明确表明在质量图像中观察到的灰质和白质之间的脂质组成差异不是由于每个解剖区域的微环境对信号强度的影响造成的假象,而是脂质组成的真实变化。