Clirim Jetishi, Erina A Balmer, Bianca M Berger, Carmen Faso, Torsten Ochsenreiter
{"title":"使用优化的 U-ExM 方案扩增蓝氏贾第鞭毛虫体内的代谢标记内细胞器和细胞骨架结构。","authors":"Clirim Jetishi, Erina A Balmer, Bianca M Berger, Carmen Faso, Torsten Ochsenreiter","doi":"10.15698/mic2024.06.825","DOIUrl":null,"url":null,"abstract":"<p><p>Understanding cellular ultrastructure is tightly bound to microscopic resolution and the ability to identify individual components at that resolution. Expansion microscopy has revolutionised this topic. Here we present and compare two protocols of ultrastructure expansion microscopy that allow for 4.5-fold mostly isotropic expansion and the use of antibodies, metabolic labelling, and DNA stains to demarcate individual regions such as the endoplasmic reticulum, the nuclei, the peripheral endocytic compartments as well as the ventral disc and the cytoskeleton in <i>Giardia lamblia</i>. We present an optimised, shortened, and modular protocol that can be swiftly adjusted to the investigators needs in this important protozoan model organism.</p>","PeriodicalId":18397,"journal":{"name":"Microbial Cell","volume":null,"pages":null},"PeriodicalIF":4.1000,"publicationDate":"2024-06-21","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11224680/pdf/","citationCount":"0","resultStr":"{\"title\":\"Expansion of metabolically labelled endocytic organelles and cytoskeletal cell structures in <i>Giardia lamblia</i> using optimised U-ExM protocols.\",\"authors\":\"Clirim Jetishi, Erina A Balmer, Bianca M Berger, Carmen Faso, Torsten Ochsenreiter\",\"doi\":\"10.15698/mic2024.06.825\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<p><p>Understanding cellular ultrastructure is tightly bound to microscopic resolution and the ability to identify individual components at that resolution. Expansion microscopy has revolutionised this topic. Here we present and compare two protocols of ultrastructure expansion microscopy that allow for 4.5-fold mostly isotropic expansion and the use of antibodies, metabolic labelling, and DNA stains to demarcate individual regions such as the endoplasmic reticulum, the nuclei, the peripheral endocytic compartments as well as the ventral disc and the cytoskeleton in <i>Giardia lamblia</i>. We present an optimised, shortened, and modular protocol that can be swiftly adjusted to the investigators needs in this important protozoan model organism.</p>\",\"PeriodicalId\":18397,\"journal\":{\"name\":\"Microbial Cell\",\"volume\":null,\"pages\":null},\"PeriodicalIF\":4.1000,\"publicationDate\":\"2024-06-21\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11224680/pdf/\",\"citationCount\":\"0\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"Microbial Cell\",\"FirstCategoryId\":\"99\",\"ListUrlMain\":\"https://doi.org/10.15698/mic2024.06.825\",\"RegionNum\":3,\"RegionCategory\":\"生物学\",\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"2024/1/1 0:00:00\",\"PubModel\":\"eCollection\",\"JCR\":\"Q2\",\"JCRName\":\"CELL BIOLOGY\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"Microbial Cell","FirstCategoryId":"99","ListUrlMain":"https://doi.org/10.15698/mic2024.06.825","RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"2024/1/1 0:00:00","PubModel":"eCollection","JCR":"Q2","JCRName":"CELL BIOLOGY","Score":null,"Total":0}
引用次数: 0
摘要
对细胞超微结构的理解与显微镜的分辨率以及在该分辨率下识别单个成分的能力密切相关。膨胀显微镜彻底改变了这一课题。在这里,我们介绍并比较了两种超微结构扩展显微镜方案,这两种方案可实现 4.5 倍的各向同性扩展,并使用抗体、代谢标记和 DNA 染色来划分单个区域,如蓝氏贾第鞭毛虫的内质网、细胞核、外周内细胞区以及腹盘和细胞骨架。我们提出了一种优化、缩短和模块化的方案,可根据研究人员的需要迅速调整,以适应这种重要的原生动物模式生物。
Expansion of metabolically labelled endocytic organelles and cytoskeletal cell structures in Giardia lamblia using optimised U-ExM protocols.
Understanding cellular ultrastructure is tightly bound to microscopic resolution and the ability to identify individual components at that resolution. Expansion microscopy has revolutionised this topic. Here we present and compare two protocols of ultrastructure expansion microscopy that allow for 4.5-fold mostly isotropic expansion and the use of antibodies, metabolic labelling, and DNA stains to demarcate individual regions such as the endoplasmic reticulum, the nuclei, the peripheral endocytic compartments as well as the ventral disc and the cytoskeleton in Giardia lamblia. We present an optimised, shortened, and modular protocol that can be swiftly adjusted to the investigators needs in this important protozoan model organism.