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Protocol for fecal microbiota transplantation: A microaerophilic approach for mice housed in a specific pathogen-free facility. 粪便微生物群移植方案：一种在特定无病原体设施中饲养的小鼠的嗜微气方法。
IF 1.3
STAR Protocols Pub Date : 2025-03-21 Epub Date: 2025-01-07 DOI: 10.1016/j.xpro.2024.103517
Shari Wouters, Hugo Moors, Mieke Verslegers, Natalie Leys, Surbhi Malhotra-Kumar, Samir Kumar-Singh, Mohamed Mysara
{"title":"Protocol for fecal microbiota transplantation: A microaerophilic approach for mice housed in a specific pathogen-free facility.","authors":"Shari Wouters, Hugo Moors, Mieke Verslegers, Natalie Leys, Surbhi Malhotra-Kumar, Samir Kumar-Singh, Mohamed Mysara","doi":"10.1016/j.xpro.2024.103517","DOIUrl":"10.1016/j.xpro.2024.103517","url":null,"abstract":"<p><p>Recently, studies have emerged exploring the potential application of fecal microbiota transplantation (FMT) in pre-clinical settings. Here, we present a protocol for FMT for mice housed in a specific pathogen-free (SPF) facility. We describe steps for sample collection, microaerophilic processing of freshly collected fecal pellets, and administration through oral gavage. We then detail procedures for the engraftment of the bacterial community. This protocol focuses on age- and gender-matched, healthy donor mice using a mobile and cost-effective alternative to an anoxic cabinet.</p>","PeriodicalId":34214,"journal":{"name":"STAR Protocols","volume":"6 1","pages":"103517"},"PeriodicalIF":1.3,"publicationDate":"2025-03-21","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11760806/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142956072","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 CRISPR-based endogenous protein tagging in mammalian cells. 基于crispr的哺乳动物细胞内源性蛋白标记方法。
IF 1.3
STAR Protocols Pub Date : 2025-03-21 Epub Date: 2025-01-20 DOI: 10.1016/j.xpro.2024.103571
Yu-Xi Xiao, Jiarun Wei, Jason Moffat
{"title":"Protocol for CRISPR-based endogenous protein tagging in mammalian cells.","authors":"Yu-Xi Xiao, Jiarun Wei, Jason Moffat","doi":"10.1016/j.xpro.2024.103571","DOIUrl":"10.1016/j.xpro.2024.103571","url":null,"abstract":"","PeriodicalId":34214,"journal":{"name":"STAR Protocols","volume":"6 1","pages":"103571"},"PeriodicalIF":1.3,"publicationDate":"2025-03-21","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11831121/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143013055","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 inducing beige adipocytes in white adipose tissue of mouse using cold exposure and CL316,243 injection. 冷暴露和CL316,243注射诱导小鼠白色脂肪组织中米色脂肪细胞的方案。
IF 1.3
STAR Protocols Pub Date : 2025-03-21 Epub Date: 2025-01-15 DOI: 10.1016/j.xpro.2024.103337
Yalan Wu, Chenguang Wang, Xiao Yu Tian
{"title":"Protocol for inducing beige adipocytes in white adipose tissue of mouse using cold exposure and CL316,243 injection.","authors":"Yalan Wu, Chenguang Wang, Xiao Yu Tian","doi":"10.1016/j.xpro.2024.103337","DOIUrl":"10.1016/j.xpro.2024.103337","url":null,"abstract":"<p><p>White adipose tissue (WAT) beiging holds significant therapeutic potential for combating obesity. Here, we present a protocol for inducing beige WAT in mice using both cold exposure and CL316,243 treatment. We describe steps for intraperitoneal injection, and subcutaneous WAT (sWAT) isolation, dissection, and fixation. We then detail procedures for histology, whole-mount immunofluorescence (IF) staining, and extracting RNA and protein. This protocol can be used for subsequent analysis to explore the mechanisms governing beige WAT induction in experimental settings, particularly the evaluation of angiogenesis. 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":"6 1","pages":"103337"},"PeriodicalIF":1.3,"publicationDate":"2025-03-21","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11783104/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143013074","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 live imaging of axonal transport in iPSC-derived iNeurons. ipsc衍生的神经元轴突运输的实时成像方案。
IF 1.3
STAR Protocols Pub Date : 2025-03-21 Epub Date: 2025-01-12 DOI: 10.1016/j.xpro.2024.103556
Dan Dou, Erika L F Holzbaur, C Alexander Boecker
{"title":"Protocol for live imaging of axonal transport in iPSC-derived iNeurons.","authors":"Dan Dou, Erika L F Holzbaur, C Alexander Boecker","doi":"10.1016/j.xpro.2024.103556","DOIUrl":"10.1016/j.xpro.2024.103556","url":null,"abstract":"<p><p>Studies of human induced pluripotent stem cell (iPSC)-derived neurons promise important insights into neurodegenerative diseases. Here, we present a protocol for live imaging of axonal transport in glutamatergic iPSC-derived neurons (iNeurons). We describe steps for the differentiation of iPSCs into iNeurons via PiggyBac-mediated neurogenin 2 (NGN2) delivery, iNeuron culture and transfection, and the acquisition and analysis of time-lapse images. Our protocol is optimized for the widely available catalog of KOLF2.1J iPSCs with mutations relevant to neurodegenerative diseases but is also applicable to other iPSC lines. For complete details on the use and execution of this protocol, please refer to Dou et al.<sup>1</sup><sup>,</sup><sup>2</sup>.</p>","PeriodicalId":34214,"journal":{"name":"STAR Protocols","volume":"6 1","pages":"103556"},"PeriodicalIF":1.3,"publicationDate":"2025-03-21","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11772938/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142980212","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 studying the superior mesenteric artery in mice using non-invasive micro-ultrasound imaging and anatomic dissection.
IF 1.3
STAR Protocols Pub Date : 2025-03-21 Epub Date: 2025-01-30 DOI: 10.1016/j.xpro.2025.103606
Haiping Chen, Yangjing Jiang, Huanhuan Huo, Guo Zhou, Min Liang, Yijie Huang, Yunwen Hu, Ziwei Zhang, Ben He, Zhaohua Cai
{"title":"Protocol for studying the superior mesenteric artery in mice using non-invasive micro-ultrasound imaging and anatomic dissection.","authors":"Haiping Chen, Yangjing Jiang, Huanhuan Huo, Guo Zhou, Min Liang, Yijie Huang, Yunwen Hu, Ziwei Zhang, Ben He, Zhaohua Cai","doi":"10.1016/j.xpro.2025.103606","DOIUrl":"10.1016/j.xpro.2025.103606","url":null,"abstract":"<p><p>The superior mesenteric artery (SMA), originating from the abdominal aorta, provides blood supply for the spleen, pancreas, intestine, and part of the colon. Here, we present a protocol for studying the SMA in the same mouse by combining non-invasive micro-ultrasound imaging and SMA dissection. We then detail procedures for tissue preparation, sectioning, and histological analysis. By allowing accurate identification of the SMA by ultrasound and anatomical dissection, this protocol can provide insights into SMA-related diseases in animal models.</p>","PeriodicalId":34214,"journal":{"name":"STAR Protocols","volume":"6 1","pages":"103606"},"PeriodicalIF":1.3,"publicationDate":"2025-03-21","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11834953/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143075811","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 the site-specific isolation of mouse endothelial cells using the modified Häutchen technique.
IF 1.3
STAR Protocols Pub Date : 2025-03-21 Epub Date: 2025-02-06 DOI: 10.1016/j.xpro.2025.103620
Alexander Brückner, Adrian Brandtner, Sarah Rieck, Bernd K Fleischmann, Daniela Wenzel
{"title":"Protocol for the site-specific isolation of mouse endothelial cells using the modified Häutchen technique.","authors":"Alexander Brückner, Adrian Brandtner, Sarah Rieck, Bernd K Fleischmann, Daniela Wenzel","doi":"10.1016/j.xpro.2025.103620","DOIUrl":"10.1016/j.xpro.2025.103620","url":null,"abstract":"<p><p>Endothelial cells (ECs) play a major role in vascular homeostasis and pathology. They show a pronounced heterogeneity between different organs as well as within the same organ. Here, we present a protocol for the rapid site-specific isolation of murine aortic ECs. We describe steps for vessel preparation and the isolation of EC layers by freezing on glass coverslips using a modified Häutchen technique. This protocol can be adapted to investigate surface cells from other tissues as well. For complete details on the use and execution of this protocol, please refer to Brückner et al.<sup>1</sup>.</p>","PeriodicalId":34214,"journal":{"name":"STAR Protocols","volume":"6 1","pages":"103620"},"PeriodicalIF":1.3,"publicationDate":"2025-03-21","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11851281/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143374801","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 protein synthesis activity in specific cell types of the testicular lumen. 鉴定睾丸腔内特定细胞类型蛋白质合成活性的方案。
IF 1.3
STAR Protocols Pub Date : 2025-03-21 Epub Date: 2025-02-28 DOI: 10.1016/j.xpro.2025.103661
Dingfeng Zou, Kai Li, Yan Lu, Wei Yan, Wei Song
{"title":"Protocol for identifying protein synthesis activity in specific cell types of the testicular lumen.","authors":"Dingfeng Zou, Kai Li, Yan Lu, Wei Yan, Wei Song","doi":"10.1016/j.xpro.2025.103661","DOIUrl":"10.1016/j.xpro.2025.103661","url":null,"abstract":"<p><p>Protein synthesis could control spermatogenic cell fate transitions. Here, we present a protocol for visualization and quantification of newly synthesized proteins by click-chemistry-based immunofluorescence within specific spermatogenic cell types in the mice testicular lumen. We detail the processes for O-propargyl-puromycin (OPP) incorporation, antibody incubation, confocal microscope imaging, and subsequent quantification methods. This protocol is not limited to spermatogenic cells and can be adapted to investigate protein synthesis in other testicular cell types and various tissue-specific cell populations. For complete details on the use and execution of this protocol, please refer to Zou et al.<sup>1</sup>.</p>","PeriodicalId":34214,"journal":{"name":"STAR Protocols","volume":"6 1","pages":"103661"},"PeriodicalIF":1.3,"publicationDate":"2025-03-21","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11919620/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143538081","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 ecotropic pseudotyped lentiviral transduction of "murinized" human cells while decreasing risk of infection for laboratory personnel.
IF 1.3
STAR Protocols Pub Date : 2025-03-21 DOI: 10.1016/j.xpro.2025.103708
Sandra L K Weber, Thorsten Heinzel, Aishwarya Iyer-Bierhoff
{"title":"Protocol for ecotropic pseudotyped lentiviral transduction of \"murinized\" human cells while decreasing risk of infection for laboratory personnel.","authors":"Sandra L K Weber, Thorsten Heinzel, Aishwarya Iyer-Bierhoff","doi":"10.1016/j.xpro.2025.103708","DOIUrl":"https://doi.org/10.1016/j.xpro.2025.103708","url":null,"abstract":"<p><p>Lentiviral systems of transduction are valuable tools to stably express transgenes of interest in human cell lines. However, their use risks infection of the lab workers. Here, we present a protocol for lentiviral vector system pseudotyping with the murine ecotropic envelope. We detail steps for producing ecotropic pseudotyped lentiviral particles and transiently expressing the Slc7a1 receptor for murine leukemia virus in target human cells. We then describe procedures for puromycin selection and the characterization of stable cell lines for transgene expression. For complete details on the use and execution of this protocol, please refer to Iyer-Bierhoff et al.<sup>1</sup>.</p>","PeriodicalId":34214,"journal":{"name":"STAR Protocols","volume":"6 2","pages":"103708"},"PeriodicalIF":1.3,"publicationDate":"2025-03-21","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143693628","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
Protocol to recover single-cell gene expression profiles from spatial transcriptomics data using cluster computing. 利用集群计算从空间转录组学数据中恢复单细胞基因表达谱的方案。
IF 1.3
STAR Protocols Pub Date : 2025-03-21 Epub Date: 2025-02-12 DOI: 10.1016/j.xpro.2025.103636
Young Je Lee, Hao Chen, Jose Lugo-Martinez
{"title":"Protocol to recover single-cell gene expression profiles from spatial transcriptomics data using cluster computing.","authors":"Young Je Lee, Hao Chen, Jose Lugo-Martinez","doi":"10.1016/j.xpro.2025.103636","DOIUrl":"10.1016/j.xpro.2025.103636","url":null,"abstract":"<p><p>Many widely used spatial transcriptomics technologies, such as Visium, capture data at multicellular resolution, precluding single-cell analysis. Here, we present scResolve, a computational protocol to recover single-cell gene expression profiles from low-resolution spatial transcriptomics data. We describe steps for computational environment setup and preparing data and formatting. We then detail procedures for running super-resolution inference and cell segmentation modules. scResolve runs in a cluster environment, leveraging parallel computing to accelerate data processing and deliver faster results. For complete details on the use and execution of this protocol, please refer to Chen et al.<sup>1</sup>.</p>","PeriodicalId":34214,"journal":{"name":"STAR Protocols","volume":"6 1","pages":"103636"},"PeriodicalIF":1.3,"publicationDate":"2025-03-21","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11969410/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143415415","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 bacterial filtration efficiency evaluations of facemask materials using viable bacteria-laden aerosol droplets. 使用含活菌气溶胶液滴对口罩材料进行细菌过滤效率评估的规程。
IF 1.3
STAR Protocols Pub Date : 2025-03-21 Epub Date: 2024-12-19 DOI: 10.1016/j.xpro.2024.103506
Mathura Thirugnanasampanthar, Ekaterina Kvitka, Mellissa Gomez, Kyle Jackson, Fereshteh Bayat, Rod G Rhem, Myrna B Dolovich, Zeinab Hosseinidoust
{"title":"Protocol for bacterial filtration efficiency evaluations of facemask materials using viable bacteria-laden aerosol droplets.","authors":"Mathura Thirugnanasampanthar, Ekaterina Kvitka, Mellissa Gomez, Kyle Jackson, Fereshteh Bayat, Rod G Rhem, Myrna B Dolovich, Zeinab Hosseinidoust","doi":"10.1016/j.xpro.2024.103506","DOIUrl":"10.1016/j.xpro.2024.103506","url":null,"abstract":"<p><p>Here, we present a protocol for determining the bacterial filtration efficiencies of facemask materials according to the American Society for Testing and Materials (ASTM) standard F2101-19. We describe steps for reproducibly generating, collecting, and enumerating viable bacteria-laden aerosols containing the bacterial pathogen Staphylococcus aureus. The optimized operating parameters generate 1,700 to 3,000 viable bacteria-laden aerosol droplets between 2.7 and 3.3 μm. For complete details on the use and execution of this protocol, please refer to Thirugnanasampanthar et al.<sup>1</sup>.</p>","PeriodicalId":34214,"journal":{"name":"STAR Protocols","volume":"6 1","pages":"103506"},"PeriodicalIF":1.3,"publicationDate":"2025-03-21","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11729654/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142872842","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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