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免疫标记在免疫质谱成像过程中扰乱了内源和抗体结合的元素度

Monique G Mello¹, Mika T Westerhausen¹, Thomas E Lockwood¹

¹Hyphenated Mass Spectrometry Laboratory, Faculty of Science, University of Technology Sydney, P.O. Box 123, Broadway, NSW, 2007, Australia.

Analytical and bioanalytical chemistry

|October 6, 2023

概括

免疫质谱成像 (LA-ICP-MS) 显示,抗体标记会改变组织中的内源金属水平 (Cu, Zn). 有机安装介质比水性介质更好地保存元素标签.

科学领域:

生物医学成像学生物医学成像学
分析化学是一种分析化学.
材料科学是一种材料科学.

背景情况:

免疫质谱成像 (LA-ICP-MS) 能够对生物分子和内源元素进行多重量化量化.
样品的处理,包括免疫标记和盖子的滑落,可以影响组织部分的元素组成.
了解这些影响对于准确地进行生物分子和金属的空间分析至关重要.

研究的目的:

调查免疫标记和覆盖粘贴对内源金属度 (Cu, Zn) 和合抗体信号的影响.
评估不同安装介质对元素标签完整性的影响.
为优化免疫质谱成像中的样本准备提供指导.

主要方法:

用冷固定的肌肉部分进行分析.
用Gd结合抗素抗体进行免疫标记.
使用LA-ICP-MS量化了Gd,Cu和Zn的元素度.
对比了有机和水性安装介质对Gd信号的影响.

主要成果:

主要抗体潜伏降低了 Zn 和增加了 Cu 度, Zn 从细胞质再分配到细胞膜.
在抗体化后,Cu主要局限于细胞膜.
Gd结合抗体信号没有受到有机安装介质的显著影响.
水性安装介质显著降低了Gd信号.

结论:

免疫标记的步骤改变了内源元素度和组织部分的分布.
建议分开部分用于内源元素和生物分子的定量分析.
有机安装介质是水性介质的首选,以保持基本标签完整性,在覆盖接用于补充成像.

关键词:

内生元素是内生元素.免疫质谱仪成像成像在LA-ICP-MS成像中使用.

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