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果蝇微小RNA-14模型中脂质的飞行时间二次离子质谱成像中,解冻装片与使用导电胶带进行样品制备的比较。

Comparison between thaw-mounting and use of conductive tape for sample preparation in ToF-SIMS imaging of lipids in Drosophila microRNA-14 model.

作者信息

Le Minh Uyen Thi, Son Jin Gyeong, Shon Hyun Kyoung, Park Jeong Hyang, Lee Sung Bae, Lee Tae Geol

机构信息

Department of Nano Science, University of Science and Technology (UST), Daejeon 34113, South Korea and Center for Nano-Bio Measurement, Korea Research Institute of Standards and Science (KRISS), Daejeon 34113, South Korea.

Center for Nano-Bio Measurement, Korea Research Institute of Standards and Science (KRISS), Daejeon 34113, South Korea.

出版信息

Biointerphases. 2018 Mar 30;13(3):03B414. doi: 10.1116/1.5019597.

Abstract

Time-of-flight secondary ion mass spectrometry (ToF-SIMS) imaging elucidates molecular distributions in tissue sections, providing useful information about the metabolic pathways linked to diseases. However, delocalization of the analytes and inadequate tissue adherence during sample preparation are among some of the unfortunate phenomena associated with this technique due to their role in the reduction of the quality, reliability, and spatial resolution of the ToF-SIMS images. For these reasons, ToF-SIMS imaging requires a more rigorous sample preparation method in order to preserve the natural state of the tissues. The traditional thaw-mounting method is particularly vulnerable to altered distributions of the analytes due to thermal effects, as well as to tissue shrinkage. In the present study, the authors made comparisons of different tissue mounting methods, including the thaw-mounting method. The authors used conductive tape as the tissue-mounting material on the substrate because it does not require heat from the finger for the tissue section to adhere to the substrate and can reduce charge accumulation during data acquisition. With the conductive-tape sampling method, they were able to acquire reproducible tissue sections and high-quality images without redistribution of the molecules. Also, the authors were successful in preserving the natural states and chemical distributions of the different components of fat metabolites such as diacylglycerol and fatty acids by using the tape-supported sampling in microRNA-14 (miR-14) deleted Drosophila models. The method highlighted here shows an improvement in the accuracy of mass spectrometric imaging of tissue samples.

摘要

飞行时间二次离子质谱(ToF-SIMS)成像可阐明组织切片中的分子分布,提供有关与疾病相关代谢途径的有用信息。然而,由于分析物的离域化以及样品制备过程中组织附着不充分等问题,会导致ToF-SIMS图像的质量、可靠性和空间分辨率降低,这些都是与该技术相关的不幸现象。由于这些原因,ToF-SIMS成像需要更严格的样品制备方法,以保持组织的自然状态。传统的解冻固定方法特别容易受到热效应导致的分析物分布改变以及组织收缩的影响。在本研究中,作者对包括解冻固定方法在内的不同组织固定方法进行了比较。作者使用导电胶带作为基底上的组织固定材料,因为它不需要手指加热就能使组织切片附着在基底上,并且可以减少数据采集过程中的电荷积累。通过导电胶带采样方法,他们能够获得可重复的组织切片和高质量图像,而不会使分子重新分布。此外,作者通过在缺失微小RNA-14(miR-14)的果蝇模型中使用胶带支撑采样,成功地保留了脂肪代谢物(如二酰基甘油和脂肪酸)不同成分的自然状态和化学分布。这里强调的方法显示了组织样品质谱成像准确性的提高。

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