The spatial resolution of microdissection‐based analytical methods to detect ocular lens glucose uptake, transport and metabolism are poor, whereas the multiplexing capability of fluorescence microscopy‐based approaches to simultaneously detect multiple glucose metabolites is limited in comparison with mass spectrometry‐based methods. To better understand lens glucose transport and metabolism, a more highly spatially resolved technique that maintains the fragile ocular lens tissue is required. In this study, a sample preparation method for matrix‐assisted laser desorption/ionisation imaging mass spectrometry (MALDI IMS) analysis of ocular lens glucose uptake and metabolism has been evaluated and optimised. Matrix choice, tissue preparation and normalisation strategy were determined using negative ion mode MALDI‐Fourier transform‐ion cyclotron resonance MS of bovine lens tissue and validation performed using gas chromatography‐MS. An internal standard was applied concurrently with N‐(1‐naphthyl)ethylenediamine dihydrochloride (NEDC) matrix to limit cracking of the fresh frozen lens tissue sections. MALDI IMS data were collected at a variety of spatial resolutions to detect both endogenous lens metabolites and stable isotopically labelled glucose introduced by ex vivo lens culture. Using this approach, initial steps in important metabolic processes that are linked to diabetic cataract formation were spatially mapped in the bovine lens. In the future, this method can be applied to study the dynamics of glucose uptake, transport and metabolic flux to aid in the study of diabetic lens cataract pathophysiology.

中文翻译：

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绘制正常牛眼中葡萄糖代谢产物的图：基质辅助激光解吸/电离成像质谱法的评估和优化

基于显微解剖的分析方法检测眼镜葡萄糖摄取，转运和新陈代谢的空间分辨率较差，而与基于质谱的方法相比，基于荧光显微镜的方法同时检测多种葡萄糖代谢产物的多路复用能力受到限制。为了更好地了解晶状体葡萄糖的运输和代谢，需要一种在空间上具有更高分辨力的技术来维持脆弱的眼晶状体组织。在这项研究中，已经评估和优化了用于晶状体葡萄糖摄取和代谢的基质辅助激光解吸/电离成像质谱（MALDI IMS）分析的样品制备方法。矩阵选择 使用负离子模式的牛晶状体组织MALDI-Fourier变换离子回旋共振MS确定组织的制备和标准化策略，并使用气相色谱MS进行验证。内部标准与N-（1-萘基）乙二胺二盐酸盐（NEDC）基质可限制新鲜冷冻晶状体组织切片的破裂。在各种空间分辨率下收集MALDI IMS数据，以检测通过离体晶状体培养引入的内源性晶状体代谢产物和稳定的同位素标记的葡萄糖。使用这种方法，将与糖尿病性白内障形成有关的重要代谢过程的初始步骤在牛眼晶状体上进行了空间定位。将来，该方法可用于研究葡萄糖摄取，转运和代谢通量的动力学，以帮助研究糖尿病性晶状体白内障的病理生理学。