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Protocol for measuring fatty acid absorption and intracellular lipid accumulation in intestinal cells and organoids. 测定肠细胞和类器官中脂肪酸吸收和细胞内脂质积累的方案。
IF 1.4
STAR Protocols Pub Date : 2026-09-23 DOI: 10.1016/j.xpro.2026.104853
Jiaqi Wu, Yu Gu, Shuangshuang Liu, Shupei Wang, Li-Hao Huang
{"title":"Protocol for measuring fatty acid absorption and intracellular lipid accumulation in intestinal cells and organoids.","authors":"Jiaqi Wu, Yu Gu, Shuangshuang Liu, Shupei Wang, Li-Hao Huang","doi":"10.1016/j.xpro.2026.104853","DOIUrl":"10.1016/j.xpro.2026.104853","url":null,"abstract":"<p><p>Quantitative assessment of intestinal epithelial cells is essential for studies of metabolism and disease. Here, we present a protocol to quantify fatty acid uptake and lipid accumulation in intestinal epithelial cell lines and primary intestinal organoids using a flow cytometry-based approach with BODIPY FL C<sub>16</sub>. We describe steps for assessing fatty acid uptake and accumulation in intestinal epithelial cell lines and intestinal organoids. This rapid and reproducible technique can also be adapted to investigate fatty acid metabolism in other cell types. For complete details on the use and execution of this protocol, please refer to Wu et al.<sup>1</sup>.</p>","PeriodicalId":34214,"journal":{"name":"STAR Protocols","volume":"7 4","pages":"104853"},"PeriodicalIF":1.4,"publicationDate":"2026-09-23","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13634375/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"149031600","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
Protocol for measuring above-threshold ionization spectra driven by few-cycle laser pulses in frustrated tunneling ionization regime. 在受挫隧道电离状态下测量由少周期激光脉冲驱动的阈值以上电离光谱的方法。
IF 1.4
STAR Protocols Pub Date : 2026-09-23 DOI: 10.1016/j.xpro.2026.104845
Lifeng Wang, Boning Wei, Bing Yu, Wei Zhang, Peng Ye
{"title":"Protocol for measuring above-threshold ionization spectra driven by few-cycle laser pulses in frustrated tunneling ionization regime.","authors":"Lifeng Wang, Boning Wei, Bing Yu, Wei Zhang, Peng Ye","doi":"10.1016/j.xpro.2026.104845","DOIUrl":"10.1016/j.xpro.2026.104845","url":null,"abstract":"<p><p>Above-threshold ionization (ATI) spectra in the frustrated tunneling ionization (FTI) regime demonstrate control of bound-state populations via the carrier-envelope phase (CEP). Here, we present a protocol for acquiring ATI spectra from argon gas driven by few-cycle laser pulses. We describe steps of few-cycle pulse generation, CEP stabilization and control, ATI spectral measurement, and data analysis. This approach enables potential applications in isolated attosecond pulse characterization. For complete details on the use and execution of this protocol, please refer to Wang et al.<sup>1</sup>.</p>","PeriodicalId":34214,"journal":{"name":"STAR Protocols","volume":"7 4","pages":"104845"},"PeriodicalIF":1.4,"publicationDate":"2026-09-23","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13632067/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"149031578","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
Protocol for conditional mapping of proximal interactomes of dimeric protein complexes using a BioID-based complementation approach. 使用基于bioid的互补方法对二聚体蛋白复合物的近端相互作用组进行条件映射的协议。
IF 1.4
STAR Protocols Pub Date : 2026-09-22 DOI: 10.1016/j.xpro.2026.104878
Abishankari Rajkumar, Antoine Gaudreau-Lapierre, Curtis L F Anthony, Laura Trinkle-Mulcahy
{"title":"Protocol for conditional mapping of proximal interactomes of dimeric protein complexes using a BioID-based complementation approach.","authors":"Abishankari Rajkumar, Antoine Gaudreau-Lapierre, Curtis L F Anthony, Laura Trinkle-Mulcahy","doi":"10.1016/j.xpro.2026.104878","DOIUrl":"10.1016/j.xpro.2026.104878","url":null,"abstract":"","PeriodicalId":34214,"journal":{"name":"STAR Protocols","volume":"7 4","pages":"104878"},"PeriodicalIF":1.4,"publicationDate":"2026-09-22","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13632053/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"149026125","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
Protocol for repeated Plasmodium chabaudi infection in a murine malaria model. 小鼠疟疾模型中反复感染沙伯地疟原虫的实验。
IF 1.4
STAR Protocols Pub Date : 2026-09-21 DOI: 10.1016/j.xpro.2026.104850
Federica Bruno, Jean Langhorne, Cristina Lo Celso
{"title":"Protocol for repeated Plasmodium chabaudi infection in a murine malaria model.","authors":"Federica Bruno, Jean Langhorne, Cristina Lo Celso","doi":"10.1016/j.xpro.2026.104850","DOIUrl":"10.1016/j.xpro.2026.104850","url":null,"abstract":"<p><p>In malaria-endemic regions, individuals experience repeated malaria infection throughout life. Here, we present a protocol to investigate the immunological mechanisms triggered by repeated exposure using a murine model of sequential infections with the rodent parasite Plasmodium chabaudi chabaudi. We provide a detailed and standardized series of steps covering primary infection, natural recovery without treatment, and subsequent secondary infection. This approach offers a robust framework to investigate the effects and long-term consequences of repeated malaria infections on the host. For complete details on use and execution of this protocol, please refer to Stephen et al.,<sup>1</sup> Pérez-Mazliah et al.,<sup>2</sup> and Achtman et al.<sup>3</sup>.</p>","PeriodicalId":34214,"journal":{"name":"STAR Protocols","volume":"7 4","pages":"104850"},"PeriodicalIF":1.4,"publicationDate":"2026-09-21","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13629323/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"149018225","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
Protocol for spatially restricted CreERT2-mediated recombination in mouse brain-infiltrating B cells using intracerebral 4-hydroxy-tamoxifen. 脑内4-羟基他莫昔芬对小鼠脑浸润性B细胞空间限制性creert2介导重组的研究
IF 1.4
STAR Protocols Pub Date : 2026-09-21 DOI: 10.1016/j.xpro.2026.104856
Mika Schneider, Georgios Perdikaris, Tobias Derfuss, Nicholas S R Sanderson
{"title":"Protocol for spatially restricted CreERT2-mediated recombination in mouse brain-infiltrating B cells using intracerebral 4-hydroxy-tamoxifen.","authors":"Mika Schneider, Georgios Perdikaris, Tobias Derfuss, Nicholas S R Sanderson","doi":"10.1016/j.xpro.2026.104856","DOIUrl":"10.1016/j.xpro.2026.104856","url":null,"abstract":"<p><p>Tamoxifen-dependent, cre-recombinase-mediated techniques enable precise temporal control of gene expression, but do not always provide sufficient anatomical control. Here, we present a protocol to restrict gene expression using local injection of 4-hydroxy-tamoxifen in bone wax. As an example, we describe the induction of the viral protein LMP1 and the fluorescent protein tdTomato only in brain-infiltrating B cells. We describe steps for intracerebral virus injection and 4-hydroxy-tamoxifen administration in mice. For complete details on the use and execution of this protocol, please refer to Kim et al.<sup>1</sup>.</p>","PeriodicalId":34214,"journal":{"name":"STAR Protocols","volume":"7 4","pages":"104856"},"PeriodicalIF":1.4,"publicationDate":"2026-09-21","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13629331/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"149017489","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
Protocol for paired isolation and characterization of migrating cancer cell subpopulations in 3D hypoxic microenvironments. 在三维缺氧微环境中对迁移癌细胞亚群进行配对分离和表征的方案。
IF 1.4
STAR Protocols Pub Date : 2026-09-19 DOI: 10.1016/j.xpro.2026.104847
Luana Schito, Sergio Rey-Keim
{"title":"Protocol for paired isolation and characterization of migrating cancer cell subpopulations in 3D hypoxic microenvironments.","authors":"Luana Schito, Sergio Rey-Keim","doi":"10.1016/j.xpro.2026.104847","DOIUrl":"10.1016/j.xpro.2026.104847","url":null,"abstract":"<p><p>Paired isolation and characterization in 3D hypoxic microenvironments (HyPIC-3D) is a platform for delineating migratory heterogeneity among cancer cell subpopulations within 3D hypoxic tumor microenvironments. Here, we present the protocol for implementing HyPIC-3D using standard laboratory and computational resources. We describe the steps to perform migration assays with cancer spheroids, collect paired non-migrated and migrated subpopulations, and characterize their heterogeneous phenotypes through quantitative imaging, molecular, and cellular analyses. This protocol has potential for applications in diverse 3D migration contexts. For complete details on the use and execution of this protocol, please refer to Schito and Rey-Keim<sup>1</sup>.</p>","PeriodicalId":34214,"journal":{"name":"STAR Protocols","volume":"7 4","pages":"104847"},"PeriodicalIF":1.4,"publicationDate":"2026-09-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13627921/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"149009003","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
Protocol for cyclic DNA-damaging treatments to generate high mutational burden in the human A549 cell line. 在人A549细胞系中产生高突变负担的循环dna损伤处理方案。
IF 1.4
STAR Protocols Pub Date : 2026-09-18 DOI: 10.1016/j.xpro.2026.104842
Gergely Imre, Szilvia Juhász
{"title":"Protocol for cyclic DNA-damaging treatments to generate high mutational burden in the human A549 cell line.","authors":"Gergely Imre, Szilvia Juhász","doi":"10.1016/j.xpro.2026.104842","DOIUrl":"10.1016/j.xpro.2026.104842","url":null,"abstract":"<p><p>Here, we present a protocol that describes iterative exposure of the human lung cancer cell line A549 to DNA-damaging agents to progressively increase mutational burden. We describe steps for controlled cytotoxic dosing, metabolic activation, and repeated treatment cycles of up to 70 iterations, followed by clonal isolation. This approach enables the generation of highly mutated cell populations suitable for downstream genomic analyses. For complete details on the use and execution of this protocol, please refer to Juhász et al.<sup>1</sup>.</p>","PeriodicalId":34214,"journal":{"name":"STAR Protocols","volume":"7 4","pages":"104842"},"PeriodicalIF":1.4,"publicationDate":"2026-09-18","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13602311/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"149002721","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
Protocol for identifying cellular and molecular signatures from spatial profiling data using micro-tissue analysis. 使用显微组织分析从空间剖面数据中识别细胞和分子特征的协议。
IF 1.4
STAR Protocols Pub Date : 2026-09-18 DOI: 10.1016/j.xpro.2026.104825
Huiqian Hu, Alphonsus H C Ng, Yue Lu
{"title":"Protocol for identifying cellular and molecular signatures from spatial profiling data using micro-tissue analysis.","authors":"Huiqian Hu, Alphonsus H C Ng, Yue Lu","doi":"10.1016/j.xpro.2026.104825","DOIUrl":"10.1016/j.xpro.2026.104825","url":null,"abstract":"<p><p>Spatial tissue profiling measures hundreds to thousands of biomolecules across tissue regions, but most analyses examine each feature one at a time. Here, we present a protocol for micro-tissue analysis using partial least-squares modeling. We describe steps for stratifying regions by microenvironment, building multivariate models to identify which features collectively drive a biological outcome, and interpreting the results. We detail four worked examples using two published GeoMx digital spatial profiler (DSP) datasets from human tumor and mucosal tissues. For complete details on the use and execution of this protocol, please refer to Lu et al.<sup>1</sup> and Ng et al.<sup>2</sup>.</p>","PeriodicalId":34214,"journal":{"name":"STAR Protocols","volume":"7 4","pages":"104825"},"PeriodicalIF":1.4,"publicationDate":"2026-09-18","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13602308/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"149002749","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
Protocol for designing and evaluating antibody-drug conjugates to promote endocytosis and lysosomal trafficking. 设计和评估促进内吞作用和溶酶体运输的抗体-药物偶联物的方案。
IF 1.4
STAR Protocols Pub Date : 2026-09-17 DOI: 10.1016/j.xpro.2026.104846
Xinlei Zhuang, Li Lin, Liqiang Pan
{"title":"Protocol for designing and evaluating antibody-drug conjugates to promote endocytosis and lysosomal trafficking.","authors":"Xinlei Zhuang, Li Lin, Liqiang Pan","doi":"10.1016/j.xpro.2026.104846","DOIUrl":"10.1016/j.xpro.2026.104846","url":null,"abstract":"<p><p>Receptor endocytosis and lysosomal delivery are critical for efficient intracellular delivery of antibody-drug conjugates (ADCs), yet many cancer targets exhibit limited internalization. Here, we present a protocol for engineering and evaluating ADCs to induce receptor ubiquitination, thereby promoting receptor uptake and transport toward lysosomal compartments. We describe steps for designing antibodies fused with E3 ligase-binding modules, drug conjugation, and cellular assays to quantify ubiquitination-dependent internalization and lysosomal accumulation in cancer cell models. For complete details on the use and execution of this protocol, please refer to Zhuang et al.<sup>1</sup>.</p>","PeriodicalId":34214,"journal":{"name":"STAR Protocols","volume":"7 4","pages":"104846"},"PeriodicalIF":1.4,"publicationDate":"2026-09-17","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13601368/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148991438","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
Protocol for assessing protein expression levels and localization during fission yeast cytokinesis. 评估分裂酵母细胞分裂过程中蛋白质表达水平和定位的方案。
IF 1.4
STAR Protocols Pub Date : 2026-09-17 DOI: 10.1016/j.xpro.2026.104839
Jack R Gregory, Yanfang Ye, Davinder Singh, Aysha H Osmani, Jian-Qiu Wu
{"title":"Protocol for assessing protein expression levels and localization during fission yeast cytokinesis.","authors":"Jack R Gregory, Yanfang Ye, Davinder Singh, Aysha H Osmani, Jian-Qiu Wu","doi":"10.1016/j.xpro.2026.104839","DOIUrl":"10.1016/j.xpro.2026.104839","url":null,"abstract":"<p><p>The fission yeast Schizosaccharomyces pombe is a genetically tractable model that shares mechanisms for cytokinesis with higher eukaryotes. Here, we present a protocol for analyzing fission yeast cytokinesis proteins. We include steps for gene targeting by homologous recombination, functionality tests by genetic interactions, and imaging by high-spatiotemporal resolution microscopy. Next, we describe procedures for image analyses using Fiji, protein interactions by co-immunoprecipitation, and measuring protein levels by Western blotting. These techniques are transferable to other subcellular locations and model organisms. For complete details on the use and execution of this protocol, please refer to Ye et al.<sup>1</sup>.</p>","PeriodicalId":34214,"journal":{"name":"STAR Protocols","volume":"7 4","pages":"104839"},"PeriodicalIF":1.4,"publicationDate":"2026-09-17","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13602322/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"149002732","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
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