Controlled Humidity Levels for Fine Spatial Detail Information in Enzyme-Assisted N-Glycan MALDI MSI,ACS Environmental Au

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Controlled Humidity Levels for Fine Spatial Detail Information in Enzyme-Assisted N-Glycan MALDI MSI
ACS Environmental Au ( IF 6.7 ) Pub Date : 2022-07-08 , DOI: 10.1021/jasms.2c00120
Dušan Veličković ₁ , Kumar Sharma ₂ , Theodore Alexandrov ₃ , Jeffrey B. Hodgin ₄ , Christopher R. Anderton ₁

Affiliation

Investigation of the spatial distribution of N-glycans in tissue specimens has emerged as a powerful tool in clinical research, in part, because altered N-glycans are often a hallmark of disease progression. Mass spectrometry imaging of N-glycans relies on peptide N-glycanase spraying and tissue incubation for efficient in situ release of N-glycans from their carrier proteins. Unstandardized and uncontrolled incubation steps often cause significant delocalization of released N-glycans, resulting in the inability to link given N-glycan composition to a specific microanatomical region in the tissue. Herein, we optimized the incubation step to provide accurate and sensitive MALDI-MSI of N-glycans. Specifically, we tested saturated solutions of various salts that maintain constant relative humidity in the incubation chamber. We showed that the best performance was achieved using a saturated solution of KNO₃ that maintains an 89% RH. Under these conditions, near maximal sensitivity was achieved with the minutest ion delocalization, which we demonstrated at a 35 μm spatial resolution, where we observed six distinct spatial patterns that colocalize to distinct microanatomical compartments in a kidney nephrectomy tissue section.

中文翻译：

酶辅助 N-聚糖 MALDI MSI 中精细空间细节信息的受控湿度水平

研究组织标本中N-聚糖的空间分布已成为临床研究中的有力工具，部分原因是改变的N-聚糖通常是疾病进展的标志。N-聚糖的质谱成像依赖于肽N-聚糖酶喷雾和组织孵育，以从其载体蛋白中有效地原位释放N-聚糖。未标准化和不受控制的孵育步骤通常会导致释放的N -聚糖显着离域，导致无法连接给定的N-组织中特定微解剖区域的聚糖成分。在此，我们优化了孵育步骤以提供准确和灵敏的N-聚糖 MALDI-MSI 。具体来说，我们测试了在孵化室中保持恒定相对湿度的各种盐的饱和溶液。我们表明，使用保持 89% RH的 KNO _{3饱和溶液可获得最佳性能。}在这些条件下，通过最微小的离子离域实现了接近最大的灵敏度，我们以 35 μm 的空间分辨率证明了这一点，我们观察到六个不同的空间模式，它们共定位到肾切除组织切片中的不同显微解剖隔室。

更新日期：2022-07-08

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