On-line reverse isotope dilution analysis for spatial quantification of elemental labels used in immunohistochemical assisted imaging mass spectrometry via LA-ICP-MS
On-line reverse isotope dilution analysis for spatial quantification of elemental labels used in immunohistochemical assisted imaging mass spectrometry via LA-ICP-MS
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DOI：
10.1039/c8ja00324f
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发表时间：
2019-02-01
影响因子：
3.4
通讯作者：
Doble, Philip A.
中科院分区：
文献类型：
--
作者：
Clases, David;de Vega, Raquel Gonzalez;Doble, Philip A.
我们提出了一种新的在线同位素稀释分析（IDA）的方法，用于定量同位素富集的金属标记物在免疫组织化学辅助成像质谱。该技术进展最近报道的在线IDA的激光烧蚀-电感耦合等离子体质谱（LA-ICP-MS），通过进行在线反向IDA的同位素富集的元素通常用作标记抗体。这种方法允许生物分子的相对定量，并且上级于具有长采集时间的其他方法，例如三维LA-ICP-MS，或者当需要更高的样品通量时。作为概念证明，用Yb同位素标记抗酪氨酸羟化酶，并在小鼠脑的两个平行冷冻切片上孵育。IDA依赖性参数通过消融基质匹配的标准品来确定。IDA定量与外部校准进行了比较，IDA是一种更耐用的方法，不受血浆漂移或自发血浆波动引起的灵敏度变化的影响。
We present a novel on-line isotope dilution analysis (IDA) approach for the quantification of isotopically enriched metal labels used in immunohistochemical assisted imaging mass spectrometry. This technique advances recently reported on-line IDA for laser ablation-inductively coupled plasma mass spectrometry (LA-ICP-MS) by performing on-line reverse IDA of isotopically enriched elements commonly used as labels for antibodies. This approach allows relative quantification of biomolecules and is superior to other methods that have long acquisition times such as three-dimensional LA-ICP-MS, or when higher sample throughput is required. As a proof of concept, anti-tyrosine hydroxylase was labelled with Yb isotopes and incubated on two parallel cryocut sections of a mouse brain. IDA dependent parameters were determined by ablation of matrix-matched standards. Quantification by IDA was compared against external calibration and was a more robust method, unaffected by sensitivity changes originating from plasma drifts or spontaneous plasma fluctuations.