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  • 文章类型: Journal Article
    具有突出的收集和转移能力的空气过滤膜的构造对于检测空气传播的病原体是高度期望的,但仍然具有挑战性。这里,通过乙醇诱导的相分离策略,直接构建了具有可调异质微纳米多孔结构的透明质酸空气过滤膜(HAFM)。由于去除效率和压降之间的理想权衡,可以通过HAFM以高性能捕获和收集空气中的病原体。通过免除样品洗脱和提取过程,过滤采样后的HAFM不仅可以直接分散在琼脂平板上进行菌落培养,还可以转化为水溶液进行离心富集,这显著减少了捕获的微生物的损害和损失。以下与ATP生物发光的组合赋予HAFM对捕获的空气传播病原体的实时定量检测功能。受益于空气病原体的高效采样和非创伤性转移,真实世界的生物气溶胶浓度可以通过基于HAFM的ATP测定来轻松评估。因此,这项工作不仅提供了一种可行的策略来制造用于高效微生物收集和富集的空气过滤膜,而且还为设计用于现场实时检测生物气溶胶的高级协议提供了启示。
    Construction of air filter membranes bearing prominent collecting and transferring capability is highly desirable for detecting airborne pathogens but remains challenging. Here, a hyaluronic acid air filter membrane (HAFM) with tunable heterogeneous micro-nano porous structures is straightforwardly constructed through the ethanol-induced phase separation strategy. Airborne pathogens can be trapped and collected by HAFM with high performance due to the ideal trade-off between removal efficiency and pressure drop. By exempting the sample elution and extraction processes, the HAFM after filtration sampling can not only directly disperse on the agar plate for colony culture but also turn to an aqueous solution for centrifugal enrichment, which significantly reduces the damage and losses of the captured microorganisms. The following combination with ATP bioluminescence endows the HAFM with a real-time quantitative detection function for the captured airborne pathogens. Benefiting from high-efficiency sampling and non-traumatic transfer of airborne pathogens, the real-world bioaerosol concentration can be facilely evaluated by the HAFM-based ATP assay. This work thus not only provides a feasible strategy to fabricate air filter membranes for efficient microbial collection and enrichment but also sheds light on designing advanced protocols for real-time detection of bioaerosols in the field.
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  • 文章类型: Historical Article
    分子工具是生态学和生物多样性科学不可或缺的一部分,并在所有生物群落中实施。大约十年前,使用和实施环境DNA(eDNA)来检测从环境样本中提取的生物多样性信号开辟了新的研究途径。最初的eDNA研究集中在理解目标物种的种群动态。此后,它的范围扩大了,在所有分类群的经过充分研究和未充分研究的生态系统中,通过元编码发现以前未记录的生物多样性。eDNA的应用迅速成为生物多样性研究的既定组成部分,和自己的研究领域。这里,我们重新审视2012年在“分子生态学”中eDNA的地标特刊中提出的关键期望,以在六个关键领域构建发展框架:(1)样本收集,(2)引物开发,(3)生物监测,(4)量化,(5)DNA在环境中的行为和(6)参考数据库的开发。我们确定了eDNA的成功,还讨论了不足和未达到的期望,突出研究重点领域,并确定意外的发展。并行,我们的回顾性研究将同行评审文献的筛选与包括学者在内的eDNA用户的调查相结合,最终用户和商业提供商,在其中,我们解决了重点研究工作以推进eDNA领域的优先领域。随着新技术进步的快速和不断增长的步伐,eDNA的未来看起来很光明,然而,成功的应用和最佳实践必须变得更加跨学科,才能充分发挥其潜力。我们的回顾给出了具体推动该领域向前发展的工具和期望。
    Molecular tools are an indispensable part of ecology and biodiversity sciences and implemented across all biomes. About a decade ago, the use and implementation of environmental DNA (eDNA) to detect biodiversity signals extracted from environmental samples opened new avenues of research. Initial eDNA research focused on understanding population dynamics of target species. Its scope thereafter broadened, uncovering previously unrecorded biodiversity via metabarcoding in both well-studied and understudied ecosystems across all taxonomic groups. The application of eDNA rapidly became an established part of biodiversity research, and a research field by its own. Here, we revisit key expectations made in a land-mark special issue on eDNA in Molecular Ecology in 2012 to frame the development in six key areas: (1) sample collection, (2) primer development, (3) biomonitoring, (4) quantification, (5) behaviour of DNA in the environment and (6) reference database development. We pinpoint the success of eDNA, yet also discuss shortfalls and expectations not met, highlighting areas of research priority and identify the unexpected developments. In parallel, our retrospective couples a screening of the peer-reviewed literature with a survey of eDNA users including academics, end-users and commercial providers, in which we address the priority areas to focus research efforts to advance the field of eDNA. With the rapid and ever-increasing pace of new technical advances, the future of eDNA looks bright, yet successful applications and best practices must become more interdisciplinary to reach its full potential. Our retrospect gives the tools and expectations towards concretely moving the field forward.
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  • 文章类型: Editorial
    生物气溶胶是一个概念,用于描述悬浮在空气中的所有生物材料,包括细菌,真菌,病毒,花粉,以及它们的衍生物,如过敏原,内毒素,霉菌毒素等.在一些研究中,初级生物气溶胶颗粒(PBAP)也被创造为指空气中完整的微生物。生物气溶胶是一门多学科的研究课题,涉及许多不同的领域,如微生物学,机械工程,空气污染,医学科学,流行病学,免疫学,生物化学,物理学,纳米技术等。自1833年查尔斯·达尔文在佛得角群岛首次在空气中发现霉菌孢子以来,生物气溶胶领域已经经历了大约200年的研究历史。近几十年来,生物气溶胶领域出现了研究热潮,从而引发了许多杰出的研究机会。在理解生物气溶胶在人类健康中的作用方面已经取得了明显的进展,大气和生态影响及其各自的技术:生物气溶胶捕获,监测和灭活。最近,来自不同领域的研究人员开始为解决生物气溶胶挑战和解决关键科学问题建立桥梁,例如,生物气溶胶传播,实时检测,室内微生物,人类生物气溶胶排放,和生物防御。为了这个努力,“生物气溶胶香山科学会议-第600届”已于夏季在北京成功举办,中国。总共邀请了47名科学家和资助机构官员,包括来自海外的领先的生物气溶胶专家,并就生物气溶胶的进展和问题进行了为期两天的广泛讨论。与会者在会议期间概述了未来的生物气溶胶方向。一些参与者也为这个生物气溶胶特刊做出了贡献。本特刊由来自八个国家的20篇生物气溶胶文章组成,包括一篇评论,并有助于生物气溶胶排放的进步,传输,对健康的影响,环境生物气溶胶,方法开发和仪器,和控制。通过这个特刊,生物气溶胶界对生物气溶胶健康风险有了更好的了解,并制定了相应的策略来应对威胁。这个特刊可能作为一个起点,不仅联系来自不同大陆的生物气溶胶科学家,而且还汇集了来自各个领域但对生物气溶胶感兴趣的人们,共同推动该领域的进一步发展。
    Bioaerosol is a concept that is used to describe all biological materials suspended in the air, including bacteria, fungi, viruses, pollen, and their derivatives such as allergens, endotoxin, mycotoxins and etc. In some studies, primary biological aerosol particle (PBAP) is also coined to refer to intact microbes in the air. Bioaerosol is a multidisciplinary research subject, involving many different fields such as microbiology, mechanical engineering, air pollution, medical science, epidemiology, immunological science, biochemistry, physics, nanotechnologies and etc. The bioaerosol field has undergone about 200 years\' research history since 1833 when mold spores were first detected in the air by Charles Darwin on the Cape Verde Islands. In recent decades, there has been a research boom in bioaerosol field, thus triggering many outstanding research opportunities. Visible progress has already been made in understanding bioaerosol roles in human health, atmospheric and ecological impacts as well as their respective technologies: bioaerosol capture, monitoring and also inactivation. Most recently, researchers from different fields start to bridge together for solving bioaerosol challenges and addressing key scientific problems, e.g., bioaerosol spread, real-time detection, indoor microbes, human bioaerosol emissions, and bio-defense. Toward this effort, a \"Bioaerosol Xiangshan Science Conference-the 600th\" has been successfully held in the summer in Beijing, China. A total of 47 scientists and funding agency officials including leading bioaerosol experts from overseas were invited and two-day long extensive discussions on bioaerosol progress and problems were carried out. Future bioaerosol directions have been outlined by the attendees during the conference. Some of the participants have also contributed to this bioaerosol special issue. This special issue consists of a total of 20 bioaerosol articles from eight countries including one review, and contributes to the advances in bioaerosol emission, transmission, health effects, ambient bioaerosols, method development and instrumentation, and control. Through this special issue, the bioaerosol community has obtained a better understanding of bioaerosol health risks and developed the corresponding strategies to confront the threats. This special issue might serve as a starting point to not only link bioaerosol scientists from different continents, but also bring together people from various fields yet with an interest in bioaerosol to collectively advance the field further.
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  • 文章类型: Journal Article
    暴露于生物气溶胶污染对人类健康具有有害影响。ATP生物发光的最新进展为定量检测生物气溶胶提供了更多机会。因为几乎所有的活性生物都能产生ATP,通过检测ATP驱动的生物发光可以容易地测量空气中微生物的量。微生物的准确评价主要依赖于以下四个关键步骤:采样和富集空气中的微生物,裂解用于ATP提取,酶促反应,和发光强度的测量。为了增强ATP生物发光的有效性,每一步都需要创新策略和持续改进。在这次审查中,综述了近年来基于ATP生物发光的空气微生物定量检测技术的研究进展,重点介绍了结合ATP生物发光改进采样装置的先进策略。同时,重点介绍了ATP生物发光测定其余三个关键步骤的优化和创新策略.目的是重新唤醒ATP生物发光的繁荣,促进其更广泛的利用,实时,和准确检测空气中的微生物。
    Exposure to bioaerosol contamination has detrimental effects on human health. Recent advances in ATP bioluminescence provide more opportunities for the quantitative detection of bioaerosols. Since almost all active organisms can produce ATP, the amount of airborne microbes can be easily measured by detecting ATP-driven bioluminescence. The accurate evaluation of microorganisms mainly relies on following the four key steps: sampling and enrichment of airborne microbes, lysis for ATP extraction, enzymatic reaction, and measurement of luminescence intensity. To enhance the effectiveness of ATP bioluminescence, each step requires innovative strategies and continuous improvement. In this review, we summarized the recent advances in the quantitative detection of airborne microbes based on ATP bioluminescence, which focuses on the advanced strategies for improving sampling devices combined with ATP bioluminescence. Meanwhile, the optimized and innovative strategies for the remaining three key steps of the ATP bioluminescence assay are highlighted. The aim is to reawaken the prosperity of ATP bioluminescence and promote its wider utilization for efficient, real-time, and accurate detection of airborne microbes.
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  • 文章类型: Journal Article
    组织内遗传异质性对健康组织和癌组织都是普遍的。它来自整个发育和体内平衡过程中体细胞突变的随机积累。通过将群体遗传学理论和基因组信息相结合,遗传异质性可以用来推断体内的组织组织和动力学。然而,许多基本数量,例如,组织特异性干细胞的动力学仍然难以精确量化。这里,我们表明,单细胞和批量测序数据提供了潜在随机过程的不同方面的信息.大量衍生的变体等位基因频谱(VAF)显示,随着年龄的增长,健康食道上皮样品中的干细胞群从生长到恒定的转变。单细胞突变负荷分布允许突变和增殖率的样本大小独立测量。与发育过程中的推论相比,成人造血干细胞的突变率更高,提示额外的增殖无关效应。此外,单细胞衍生的VAF光谱包含有关组织特异性干细胞数量的信息。在造血系统中,我们发现大约2×105个造血干细胞,如果所有干细胞对称分裂。然而,与泊松分布随机突变模型相比,单细胞突变负荷分布过度分散.仅具有恒定速率的与时间相关的突变累积模型无法生成这种模式。至少需要一个额外的随机性来源。这些过程的可能候选者可能是偶尔的干细胞分裂爆发,可能是为了应对伤害,或通过环境暴露或细胞内在变异的非恒定突变率。
    Intra-tissue genetic heterogeneity is universal to both healthy and cancerous tissues. It emerges from the stochastic accumulation of somatic mutations throughout development and homeostasis. By combining population genetics theory and genomic information, genetic heterogeneity can be exploited to infer tissue organization and dynamics in vivo. However, many basic quantities, for example the dynamics of tissue-specific stem cells remain difficult to quantify precisely. Here, we show that single-cell and bulk sequencing data inform on different aspects of the underlying stochastic processes. Bulk-derived variant allele frequency spectra (VAF) show transitions from growing to constant stem cell populations with age in samples of healthy esophagus epithelium. Single-cell mutational burden distributions allow a sample size independent measure of mutation and proliferation rates. Mutation rates in adult hematopietic stem cells are higher compared to inferences during development, suggesting additional proliferation-independent effects. Furthermore, single-cell derived VAF spectra contain information on the number of tissue-specific stem cells. In hematopiesis, we find approximately 2 × 105 HSCs, if all stem cells divide symmetrically. However, the single-cell mutational burden distribution is over-dispersed compared to a model of Poisson distributed random mutations. A time-associated model of mutation accumulation with a constant rate alone cannot generate such a pattern. At least one additional source of stochasticity would be needed. Possible candidates for these processes may be occasional bursts of stem cell divisions, potentially in response to injury, or non-constant mutation rates either through environmental exposures or cell-intrinsic variation.
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  • 文章类型: Journal Article
    植物中的天然化合物通常分布不均,确定最佳采样位置以获得最具代表性的结果在技术上具有挑战性。基质辅助激光解吸/电离质谱成像(MALDI-MSI)可以为制定采样指南提供依据。对于多汁植物样本,在MSI分析过程中确保空间分布分析结果的真实性和原位性质也需要充分考虑。在这项研究中,我们开发了一种基于保存方法的完善和可靠的MALDI-MSI方法,切片条件,辅助矩阵,和MALDI参数,以检测和可视化马斯卡林在落叶松的原位空间分布。使用液相色谱-串联质谱法验证MALDI-MSI结果。-80℃低温储存和"书签"干燥是多肉植物样品及其花卉样品的适宜储存方法,并在-20℃下使用明胶作为包埋介质切成40μm厚的切片是合适的切片方法。使用DCTB(反式-2-[3-(4-叔丁基苯基)-2-甲基-2-亚丙烯基]丙二腈)作为辅助基质,激光强度为45是MALDI参数的有利条件mescaline分析。感兴趣的区域半定量分析显示,mescaline集中在L.williamsii的表皮组织以及牙冠的分生组织中。这项研究结果不仅有助于为确定威廉姆卡林的最佳采样位置提供依据。同时也为MALDI检测前植物器官的贮藏和制备条件的优化提供了参考。
    基于多参数比较实验,获得了一种用于富含淡水的多肉植物的准确原位MSI方法。使用上述方法进行了米斯卡林的空间成像分析。根据以上结果和以前的结果,初步提出了法医学实践的抽样建议。
    Natural compounds in plants are often unevenly distributed, and determining the best sampling locations to obtain the most representative results is technically challenging. Matrix-assisted laser desorption/ionization mass spectrometry imaging (MALDI-MSI) can provide the basis for formulating sampling guideline. For a succulent plant sample, ensuring the authenticity and in situ nature of the spatial distribution analysis results during MSI analysis also needs to be thoroughly considered. In this study, we developed a well-established and reliable MALDI-MSI method based on preservation methods, slice conditions, auxiliary matrices, and MALDI parameters to detect and visualize the spatial distribution of mescaline in situ in Lophophora williamsii. The MALDI-MSI results were validated using liquid chromatography-tandem mass spectrometry. Low-temperature storage at -80°C and drying of \"bookmarks\" were the appropriate storage methods for succulent plant samples and their flower samples, and cutting into 40 μm thick sections at -20°C using gelatin as the embedding medium is the appropriate sectioning method. The use of DCTB (trans-2-[3-(4-tert-butylphenyl)-2-methyl-2-propenylidene]malononitrile) as an auxiliary matrix and a laser intensity of 45 are favourable MALDI parameter conditions for mescaline analysis. The region of interest semi-quantitative analysis revealed that mescaline is concentrated in the epidermal tissues of L. williamsii as well as in the meristematic tissues of the crown. The study findings not only help to provide a basis for determining the best sampling locations for mescaline in L. williamsii, but they also provide a reference for the optimization of storage and preparation conditions for raw plant organs before MALDI detection.
    UNASSIGNED: An accurate in situ MSI method for fresh water-rich succulent plants was obtained based on multi-parameter comparative experiments.Spatial imaging analysis of mescaline in Lophophora williamsii was performed using the above method.Based on the above results and previous results, a sampling proposal for forensic medicine practice is tentatively proposed.
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  • 文章类型: Journal Article
    信息驱动的任务中止是控制任务执行过程中安全关键系统故障风险的有效方法。我们研究了部分可观察到的安全关键系统的最佳采样和任务中止决策,其中,底层系统健康状态只能通过抽样来揭示。与以前的研究相比,我们采用部分健康信息来共同确定:(A)是否执行采样,以及(b)何时以动态方式中止任务,以便最小化抽样产生的预期总成本,任务失败,和系统故障。动态采样和任务中止策略是根据信念状态设计的,其优化模型被投入到部分可观察的马尔可夫决策过程的框架中。关于价值函数的一些结构性见解,控制极限选择,并给出了最优性存在性。通过数值实验检验了所提出的采样和中止策略的性能,在任务损失控制中,这些策略被证明优于其他启发式中止策略。
    Information-driven mission abort is an effective way to control the failure risk of safety-critical systems during mission executions. We investigate the optimal sampling and mission abort decisions of partially observable safety-critical systems, where the underlying system health state can only be revealed by sampling. In contrast to previous studies, we employ partial health information to jointly determine: (a) whether to execute sampling, and (b) when to abort the mission in a dynamic manner, so as to minimize the expected total cost incurred by sampling, mission failure, and system malfunction. Dynamic sampling and mission abort policies are devised following the belief state, whose optimization model is cast into the framework of a partially observable Markov decision process. Some structural insights with regard to the value function, control limit selection, and optimality existence are presented. The performance of the proposed sampling and abort policy is tested by numerical experiments, which are proved to outperform other heuristic abort policies in mission loss control.
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  • 文章类型: Journal Article
    通常需要通过常规分析方法对大气气溶胶进行化学分析,以执行复杂且耗时的样品制备过程。近几十年来,环境电离质谱(AI-MS)方法已被证明是简单的,快速,以及直接分析各种复杂样品的有效分析工具。在这项工作中,我们使用多孔纸过滤器对烟草烟雾进行直接吸附采样,然后取样的纸过滤器被执行纸喷雾电离(PSI)装置的发射器。制造自动采样装置以控制烟草烟雾的产生和收集。尼古丁,烟草烟雾的典型化合物,用于优化自动采样的关键条件。此外,还通过多变量分析比较了不同类型的烟草烟雾,并调查了来自不同来源的烟草烟雾的制造者。通过使用此方法,可以在几分钟内完成单个烟草样品的直接采样和分析。总的来说,我们的结果表明,PSI-MS是一个强大的工具,集成收集,提取,电离,和识别烟雾中的分析物。
    Chemical analysis of atmospheric aerosols by conventional analytical methods is usually required to perform complicated and time-consuming sample preparation processes. In recent decades, ambient ionization mass spectrometry (AI-MS) methods have been proven to be simple, rapid, and effective analytical tools for direct analysis of various complex samples. In this work, we applied porous paper filters for direct adsorptive sampling of tobacco smoke, and then the sampled paper filters were performed the emitters of the paper spray ionization (PSI) device. An auto-sampling device was made to control the generation and collection of tobacco smoke. Nicotine, the typical compound of tobacco smoke, was used to optimize the key conditions of auto-sampling. Moreover, different types of tobacco smoke were also compared with multivariate variable analysis, and the makers of tobacco smoke from different sources of tobacco smoke were investigated. By using this method, direct sampling and analysis of a single tobacco sample can be completed within minutes. Overall, our results show that PSI-MS is a powerful tool that integrates collection, extraction, ionization, and identification analytes in smoke.
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  • 文章类型: Journal Article
    越来越多的研究证明microRNAs(miRNAs)在基因表达调控中起着至关重要的作用,miRNA的不规则表达往往与多种复杂的人类疾病有关。由于通过生物学实验鉴定疾病相关miRNAs的成本高、效率低,学者们一直致力于通过计算方法预测潜在的疾病相关miRNAs。考虑到现有方法在构建负样本集时存在缺陷,我们提出了一种基于聚类的miRNA-疾病关联预测(CSMDA)抽样方法.首先,我们整合了miRNA和疾病的多个相似性信息来表示miRNA-疾病对。其次,为了避免在构建负样本集时引入潜在的正样本,我们采用了基于聚类的抽样方法.第三,我们采用了一种基于随机森林的特征选择方法来减少高维特征空间中的噪声和冗余信息。最后,我们实施了一个集成学习框架,用于通过软投票预测miRNA-疾病关联.精度,回想一下,F1分数,CSMDA的AUROC和AUPR分别达到0.9676、0.9545、0.9610、0.9928和0.9940,在五倍交叉验证下。此外,对三种癌症的个案研究表明,由CSMDA预测的前20个潜在相关miRNAs已被dbDEMC数据库或文献证实。以上结果表明CSMDA可以更准确地预测潜在的疾病相关miRNA。
    More and more studies have proved that microRNAs (miRNAs) play a critical role in gene expression regulation, and the irregular expression of miRNAs tends to be associated with a variety of complex human diseases. Because of the high cost and low efficiency of identifying disease-associated miRNAs through biological experiments, scholars have focused on predicting potential disease-associated miRNAs by computational methods. Considering that the existing methods are flawed in constructing negative sample set, we proposed a clustering-based sampling method for miRNA-disease association prediction (CSMDA). Firstly, we integrated multiple similarity information of miRNA and disease to represent miRNA-disease pairs. Secondly, we performed a clustering-based sampling method to avoid introducing potential positive samples when constructing negative sample set. Thirdly, we employed a random forest-based feature selection method to reduce noise and redundant information in the high-dimensional feature space. Finally, we implemented an ensemble learning framework for predicting miRNA-disease associations by soft voting. The Precision, Recall, F1-score, AUROC and AUPR of the CSMDA achieved 0.9676, 0.9545, 0.9610, 0.9928, and 0.9940, respectively, under five-fold cross-validation. Besides, case study on three cancers showed that the top 20 potentially associated miRNAs predicted by the CSMDA were confirmed by the dbDEMC database or literatures. The above results demonstrate that the CSMDA can predict potential disease-associated miRNAs more accurately.
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  • 文章类型: Journal Article
    从组织中直接采样脂质进行直接质谱(MS)分析可以快速分析脂质,这对生物医学应用很重要。在这项工作中,我们开发了一种多孔聚合物膜(PPM)微探针,用于直接从组织样品中高效采样脂质。PPM由聚丙烯制备,孔径为10μm,促进脂质从组织表面的渗透。PPM被涂在一根不锈钢丝上,厚度约为100μm。整个分析程序包括对组织中的脂质进行采样,洗探针,和萃取喷雾电离MS分析。通过分析小鼠脑组织样品验证了有效性。与总脂质提取方法相比,它显示出一系列脂质类别的高回收率。通过分析小鼠肝脏的组织样本进行了进一步的证明,胃,肾和腿。具有较高的物理强度和良好的化学稳定性,微探针还被证明可用于对小鼠肾脏组织样本中的脂质进行采样。通过引入光化学衍生,还开发了快速检测脂质C[键,双键]组织样品中的C异构体。最后,还开发了微探针阵列,用于从组织表面的多个部位同时采样脂质。
    Direct sampling of lipids from tissues for direct mass spectrometry (MS) analysis allows a quick profiling of lipidome, which is important for biomedical applications. In this work, we developed a polyporous polymeric membrane (PPM) microprobe for highly efficient sampling of lipids directly from tissue samples. The PPM was prepared by polypropylene with pores as large of 10 ​μm, facilitating the permeation of lipids from tissue surfaces. The PPM was coated onto a stainless steel wire with a thickness of ∼100 ​μm. The entire analysis procedure includes sampling of the lipids in tissue, washing the probe, and extraction spray ionization for MS analysis. The effectiveness was validated by analyzing mouse brain tissue samples. It showed high recoveries for a series of lipid classes in comparison with total lipid extraction method. Further demonstration was carried out with analysis of tissue samples from mouse liver, stomach, kidney and legs. With high physical strength and good chemical stability, the microprobe was also demonstrated for sampling lipids inside mouse kidney tissue samples. By incorporating a photochemical derivatization, a workflow was also developed for fast detection of lipid C[bond, double bond]C isomers in tissue samples. Finally, a microprobe array was also developed for simultaneous sampling of lipids from multiple sites on tissue surfaces.
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