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Enhancing MALDI-MSI Spatial Proteomics Analysis through Matrix Solution Acidification 通过基质溶液酸化增强MALDI-MSI空间蛋白质组学分析

IF 4.2 3区化学

Analyst Pub Date : 2025-07-25 DOI: 10.1039/d5an00446b

Gaofeng Ji, Li Yi, Kunyue Deng, Peiqi Luo, Lingpeng Zhan

{"title":"Enhancing MALDI-MSI Spatial Proteomics Analysis through Matrix Solution Acidification","authors":"Gaofeng Ji, Li Yi, Kunyue Deng, Peiqi Luo, Lingpeng Zhan","doi":"10.1039/d5an00446b","DOIUrl":"https://doi.org/10.1039/d5an00446b","url":null,"abstract":"Mass spectrometry imaging (MSI) is a powerful technique for spatial resolved proteomics analysis, offering rapid data acquisition and high lateral resolution. However, its utility is often constrained by limited proteomics coverage. In this study, we introduce a simple yet effective method to enhance the detection efficiency of matrix-assisted laser desorption/ionization (MALDI)-MSI for both intact proteins and high-mass tryptic peptides in tissues. By reformulating conventional matrices with elevated concentration of formic acid (FA), we observed that significant improvements in imaging performance, including enhanced peak detection and superior image quality. The efficacy of this approach was first validated using sinapinic acid (SA), a widely adopted matrix for intact protein imaging. Notably, the matrix acidification approach exhibited complementary benefits in tryptic peptide MSI. When using matrix α-cyano-4-hydroxycinnamic acid (CHCA) with 5% FA, we observe selective enhancement of high-mass peptides (> 1500 m/z) in MSI, along with improved image quality. Collectively, our results demonstrate the potential of this approach to improve the proteomic coverage in MSI-based spatial proteomics analysis.","PeriodicalId":63,"journal":{"name":"Analyst","volume":"48 1","pages":""},"PeriodicalIF":4.2,"publicationDate":"2025-07-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144701512","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Advanced Raman assisted metabolic labelling: an impending approach for exploring metabolites in cellular biosynthetic processes. 先进的拉曼辅助代谢标记：在细胞生物合成过程中探索代谢物的一种即将到来的方法。

IF 4.2 3区化学

Analyst Pub Date : 2025-07-25 DOI: 10.1039/d5an00586h

Madhukrishnan Murali,Kaustabh Kumar Maiti

引用次数: 0

mPCR/SERS assay for hMLH1 promoter methylation identification hMLH1启动子甲基化鉴定的mPCR/SERS分析

IF 4.2 3区化学

Analyst Pub Date : 2025-07-24 DOI: 10.1039/d5an00589b

Kazi Morshed Alom, Nana Lyu, Anastasiia Tukova, Alison Rodger, Yuling Wang

{"title":"mPCR/SERS assay for hMLH1 promoter methylation identification","authors":"Kazi Morshed Alom, Nana Lyu, Anastasiia Tukova, Alison Rodger, Yuling Wang","doi":"10.1039/d5an00589b","DOIUrl":"https://doi.org/10.1039/d5an00589b","url":null,"abstract":"Gene-specific DNA methylation is associated with the progression of various cancers. Thus accurate identification of this epigenetic alteration is of great interest. In this study, we particularly focused on the use of surface-enhanced Raman scattering (SERS) for detecting methylation in the promoter region of the hMLH1 gene found in cancer cells. A promoter segment from two different colorectal cancer cell lines, SW48 and SW480 identified as hypermethylated and unmethylated respectively, were amplified by methylation-specific PCR (mPCR). This produced two different types of amplificons with specific primers where the forward primer was labelled with completely different 5' overhang sequences, and the reverse primer was labelled with biotin. This allowed them to bind specifically with either a methylation-specific nanotag (mSERS) or an unmethylation-specific nanotag (umSERS), followed by enrichment by streptavidin magnetic beads (SMB). This resulted in the formation of a nanotag-amplicon-SMB complex that could be easily isolated from the unbound molecules. Finally, SERS analysis of the complex produced specific spectral profiles related to mSERS and umSERS nanotags, indicating the methylation status of the promoters. The mPCR/SERS assay demonstrated a limit of detection (LOD) of 3.93% for sensing DNA methylation with high specificity. Thus we believe the proposed assay can find more broad applications for methylation analysis in any DNA samples.","PeriodicalId":63,"journal":{"name":"Analyst","volume":"18 1","pages":""},"PeriodicalIF":4.2,"publicationDate":"2025-07-24","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144693654","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Use of Raman spectroscopy for the label-free discrimination of CHO cells associated with antibody expression 利用拉曼光谱对与抗体表达相关的CHO细胞进行无标记识别

IF 4.2 3区化学

Analyst Pub Date : 2025-07-24 DOI: 10.1039/d5an00476d

Yusui Sato, Yukako Senga, Hiroshi Tateno, Shinya Honda

{"title":"Use of Raman spectroscopy for the label-free discrimination of CHO cells associated with antibody expression","authors":"Yusui Sato, Yukako Senga, Hiroshi Tateno, Shinya Honda","doi":"10.1039/d5an00476d","DOIUrl":"https://doi.org/10.1039/d5an00476d","url":null,"abstract":"The performances of host cells that produce therapeutic proteins is important for the efficient production of drug substances, to minimize production costs, and to ensure competitive prices for commercial drugs in the biopharmaceutical industry. However, the selection of cells that exhibit high performances in the expression of proteins and in the construction of master cell banks is a complicated and time-consuming process. Consequently, novel methods are required to assess the expression levels of drug molecules and to select dominant cells at an early stage. In this study, Raman spectroscopy, which constitutes a label-free analytical method for obtaining molecular information derived from host cells, was used to investigate antibody expression in Chinese hamster ovary (CHO) cells. Antibody-producing and non-producing CHO cells were cultured and live cell observations were conducted using Raman spectroscopy. Multivariate analysis of the Raman spectra demonstrated different classifications of antibody-producing and non-producing CHO cells. The results of this study suggest that Raman spectroscopy has the potential to evaluate the performances of host cells for biopharmaceutical production and enhance the efficiency profiles of production lines.","PeriodicalId":63,"journal":{"name":"Analyst","volume":"123 1","pages":""},"PeriodicalIF":4.2,"publicationDate":"2025-07-24","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144693653","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Advances in metal-DOTA/DTPA complexes for mass cytometry 金属- dota /DTPA配合物在细胞计数中的研究进展

IF 4.2 3区化学

Analyst Pub Date : 2025-07-24 DOI: 10.1039/d5an00485c

Yin-Feng Wang, Wenying Wu, Guojun Han

引用次数: 0

Zirconium-Carboxylate Metal-Organic Frameworks as Stationary Phases for Chromatographic Separation 锆-羧酸盐金属-有机骨架作为色谱分离的固定相

IF 4.2 3区化学

Analyst Pub Date : 2025-07-23 DOI: 10.1039/d5an00468c

Hai-Yue Wei, Sha-Sha Meng, Jian Zhang, Mo-Han Gu, Ming Xu, Zhi-Yuan Gu

引用次数: 0

ESIPT-active fluorescent probes for ratiometric analysis of carbonate ions in aqueous media: structural modifications for enhanced selectivity and response. 用于水介质中碳酸盐离子比率分析的esipt活性荧光探针：结构修饰以增强选择性和响应。

IF 4.2 3区化学

Analyst Pub Date : 2025-07-23 DOI: 10.1039/d5an00215j

M Gayathri,Sourav Mondal,D Sriram,Nilanjan Dey

{"title":"ESIPT-active fluorescent probes for ratiometric analysis of carbonate ions in aqueous media: structural modifications for enhanced selectivity and response.","authors":"M Gayathri,Sourav Mondal,D Sriram,Nilanjan Dey","doi":"10.1039/d5an00215j","DOIUrl":"https://doi.org/10.1039/d5an00215j","url":null,"abstract":"This study focuses on the design and synthesis of two ESIPT-active fluorescent probes that differ in the position of the hydroxyl group. The isomer with the hydroxyl group at the ortho position in compound 1 exhibited a large population of the keto tautomer in equilibrium, forming more compact nanoaggregates in aqueous media. In contrast, the enol form was predominant in compound 2, which has the hydroxyl group at the terminal position, leading to significantly larger aggregate sizes compared to the first probe. Both compounds demonstrated a ratiometric color-changing response with carbonate ions, accompanied by a turn-on fluorescence response. Mechanistic studies revealed that hydrogen bonding (or possible deprotonation) induced by carbonate ions influenced the keto-enol equilibrium and conformational flexibility, which affected both the optical properties and aggregation behavior of the probes. Comparative analysis showed that compound 1 had a superior response towards carbonate ions compared to probe 2. The position of the hydroxyl group altered the binding cleft available for carbonate ions. Furthermore, the naphthyl-substituted analogue of probe 1 formed larger aggregates and exhibited a stronger response to carbonate ions than probe 1 itself. Cross-reactivity studies confirmed the high selectivity of compound 1 for carbonate ions, with minimal interference from other anions. These results motivated the use of the current system for analyzing carbonate ions in natural water samples collected from the laboratory tap and nearby ponds. The high recovery rates and low standard deviation confirmed the suitability of this system for the quantitative analysis of carbonate in real-life samples.","PeriodicalId":63,"journal":{"name":"Analyst","volume":"667 1","pages":""},"PeriodicalIF":4.2,"publicationDate":"2025-07-23","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144684303","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Portable Detection Methods for Marine Micro-nano Plastics 海洋微纳塑料的便携式检测方法

IF 4.2 3区化学

Analyst Pub Date : 2025-07-23 DOI: 10.1039/d5an00597c

Gang Chen, Jiahao Dong, Min Dai, Xiaobo Xiong, Juan Mei, Jing Pan

{"title":"Portable Detection Methods for Marine Micro-nano Plastics","authors":"Gang Chen, Jiahao Dong, Min Dai, Xiaobo Xiong, Juan Mei, Jing Pan","doi":"10.1039/d5an00597c","DOIUrl":"https://doi.org/10.1039/d5an00597c","url":null,"abstract":"Every year, millions of tons of plastic waste enter the ocean, gradually breaking into micro-nano plastics, threatening the ecosystem and human health. The detection of Marine micro-nano plastics is an important link in assessing ecological risks and guiding the implementation of governance. The traditional detection technologies rely on complex equipment and pretreatment, making it difficult to achieve rapid and on-site detection of micro-nano plastics in seawater environments. Portable detection methods, which can realize on-site and rapid distributed detection of Marine micro-nano plastics, thus attract more and more attention. Herein, recent existing portable Marine micro-nano plastics detection methods are introduced. Firstly, the conventional methods of Marine micro-nano plastics sensors and the corresponding advances in portable area are presented. The conventional methods include pyrolysis gas chromatography-mass spectrometry, infrared spectroscopy and Raman spectroscopy, Then, the novel detection methods developed in recent 5 years for portable Marine micro-nano plastics detection are discussed, such as photoluminescence spectroscopy, triboelectric nanogenerator-based self-powered sensor and electrochemical sensor. Finally, the challenges for the development of portable Marine micro-nano plastics detection methods are demonstrated for the practical application.","PeriodicalId":63,"journal":{"name":"Analyst","volume":"53 1","pages":""},"PeriodicalIF":4.2,"publicationDate":"2025-07-23","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144685018","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

HIST-DIP: Histogram Thresholding and Deep Image Priors assisted Smartphone-Based Fluorescence Microscopy Imaging histdip：直方图阈值和深度图像先验辅助基于智能手机的荧光显微镜成像

IF 4.2 3区化学

Analyst Pub Date : 2025-07-23 DOI: 10.1039/d5an00487j

Harshitha Govindaraju, Muhammad Nabeel Tahir, Umer Hassan

{"title":"HIST-DIP: Histogram Thresholding and Deep Image Priors assisted Smartphone-Based Fluorescence Microscopy Imaging","authors":"Harshitha Govindaraju, Muhammad Nabeel Tahir, Umer Hassan","doi":"10.1039/d5an00487j","DOIUrl":"https://doi.org/10.1039/d5an00487j","url":null,"abstract":"Portable fluorescence microscopes coupled with smartphones offer accessible and cost-effective point- of-care diagnostic solutions, but often produce noisy and blurry images with poor contrast. Here, we introduce HIST-DIP (HIStogram Thresholding and Deep Image Prior), an unsupervised framework for fluorescence microscopy image restoration. Histogram thresholding isolates fluorescence signals by removing background noise, while DIP refines structural details and enhances resolution without large labeled datasets. Validation results show substantial quality gains including the average Peak Signal-to-Noise Ratio (PSNR) improved from 15.59dB to 27.10dB, and the Structural Similarity Index Measure (SSIM) rose from 0.035 to 0.82. Contrast-to-noise ratio (CNR) and signal difference- to-noise ratio (SDNR) also increased significantly, indicating sharper bead outlines and reduced background interference. Unlike conventional deep learning methods, HIST-DIP needs no external training data, making it well-suited for real-time, low-cost, and point-of-care diagnostic imaging. These findings highlight the potential of HIST-DIP in enhancing the quality of smartphone-based microscopy images, while also motivating future research towards optimizing the methods for real- time on-device computations.","PeriodicalId":63,"journal":{"name":"Analyst","volume":"108 1","pages":""},"PeriodicalIF":4.2,"publicationDate":"2025-07-23","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144685019","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

A holistic approach to understanding biochemical degradation of human tissues using high resolution MALDI MS 使用高分辨率MALDI质谱分析理解人体组织生化降解的整体方法

IF 4.2 3区化学

Analyst Pub Date : 2025-07-18 DOI: 10.1039/d5an00588d

Jerika Ho, Naomi Stock, Vaughn Mangal, Shari Forbes, Theresa Stotesbury

{"title":"A holistic approach to understanding biochemical degradation of human tissues using high resolution MALDI MS","authors":"Jerika Ho, Naomi Stock, Vaughn Mangal, Shari Forbes, Theresa Stotesbury","doi":"10.1039/d5an00588d","DOIUrl":"https://doi.org/10.1039/d5an00588d","url":null,"abstract":"Estimating the postmortem interval (PMI) is crucial in medico-legal death investigations, but existing methods are highly influenced by environmental and physiological factors. This study proposes a novel biochemical strategy for PMI estimation using Matrix-Assisted Laser Desorption/Ionization Mass Spectrometry (MALDI MS) to detect molecular changes in decomposing human soft tissues. Samples from the quadriceps and shoulder were collected from 13 human donors at the Research in Experimental and Social Thanatology (REST) facility in Quebec, Canada, spanning accumulated degree days (ADD) ranging from 9.75 to 11,455.86. Tissues (14 µm) were cryosectioned, mounted on carbon tape, dried under nitrogen gas, and coated with a 2,5-dihydroxybenzoic acid matrix solution. Hematoxylin and eosin staining provided histological context for the sampled tissues. MALDI MS data were acquired using a Bruker SolariX XR Fourier-transform Ion Cyclotron Resonance MS. Van Krevelen diagrams and principal component analysis revealed decomposition-linked trends, with shifts in biochemical profiles over time. N/C and N/O ratios increased, reflecting protein and peptide degradation and microbial activity, while O/C ratios declined, likely due to the loss of oxygen-rich compounds. Mixed-effects models showed moderate associations (R² marginal = 0.06–0.49) between elemental ratios and ADD, with conditional R² values up to 0.55. Spearman’s rank correlation identified compounds significantly associated with ADD. Six candidate biomarkers were identified, with variable model fit (R² marginal = 0.003–0.80) and unresolved donor-level effects. This study demonstrates the novel application of MALDI MS for PMI biomarker discovery and contributes one of the largest datasets in forensic decomposition chemistry research.","PeriodicalId":63,"journal":{"name":"Analyst","volume":"7 1","pages":""},"PeriodicalIF":4.2,"publicationDate":"2025-07-18","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144652375","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0