Ultrastructure expansion microscopy (U-ExM) of mouse and human kidneys for analysis of subcellular structures.

التفاصيل البيبلوغرافية
العنوان:	Ultrastructure expansion microscopy (U-ExM) of mouse and human kidneys for analysis of subcellular structures.
المؤلفون:	Langner E; Department of Medicine, Washington University, St. Louis, Missouri, USA., Puapatanakul P; Department of Medicine, Washington University, St. Louis, Missouri, USA.; Division of Nephrology, Department of Medicine, Faculty of Medicine, Chulalongkorn University, Bangkok, Thailand., Pudlowski R; Department of Biology, Washington University, St. Louis, Missouri, USA., Alsabbagh DY; Department of Medicine, Washington University, St. Louis, Missouri, USA., Miner JH; Department of Medicine, Washington University, St. Louis, Missouri, USA., Horani A; Department of Pediatrics, Washington University, St. Louis, Missouri, USA., Dutcher SK; Department of Genetics, Washington University, St. Louis, Missouri, USA., Brody SL; Department of Medicine, Washington University, St. Louis, Missouri, USA., Wang JT; Division of Nephrology, Department of Medicine, Faculty of Medicine, Chulalongkorn University, Bangkok, Thailand., Suleiman HY; Department of Medicine, Washington University, St. Louis, Missouri, USA., Mahjoub MR; Department of Medicine, Washington University, St. Louis, Missouri, USA.; Department of Cell Biology and Physiology, Washington University, St. Louis, Missouri, USA.
المصدر:	Cytoskeleton (Hoboken, N.J.) [Cytoskeleton (Hoboken)] 2024 May 07. Date of Electronic Publication: 2024 May 07.
Publication Model:	Ahead of Print
نوع المنشور:	Journal Article
اللغة:	English
بيانات الدورية:	Publisher: Wiley-Liss Country of Publication: United States NLM ID: 101523844 Publication Model: Print-Electronic Cited Medium: Internet ISSN: 1949-3592 (Electronic) Linking ISSN: 19493592 NLM ISO Abbreviation: Cytoskeleton (Hoboken) Subsets: MEDLINE
أسماء مطبوعة:	Original Publication: Hoboken, NJ : Wiley-Liss, 2010-
مستخلص:	Ultrastructure expansion microscopy (U-ExM) involves the physical magnification of specimens embedded in hydrogels, which allows for super-resolution imaging of subcellular structures using a conventional diffraction-limited microscope. Methods for expansion microscopy exist for several organisms, organs, and cell types, and used to analyze cellular organelles and substructures in nanoscale resolution. Here, we describe a simple step-by-step U-ExM protocol for the expansion, immunostaining, imaging, and analysis of cytoskeletal and organellar structures in kidney tissue. We detail the critical modified steps to optimize isotropic kidney tissue expansion, and preservation of the renal cell structures of interest. We demonstrate the utility of the approach using several markers of renal cell types, centrioles, cilia, the extracellular matrix, and other cytoskeletal elements. Finally, we show that the approach works well on mouse and human kidney samples that were preserved using different fixation and embedding conditions. Overall, this protocol provides a simple and cost-effective approach to analyze both preclinical and clinical renal samples in high detail, using conventional lab supplies and standard widefield or confocal microscopy. (© 2024 Wiley Periodicals LLC.)
التعليقات:	Update of: bioRxiv. 2024 Feb 17;:. (PMID: 38405695)
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معلومات مُعتمدة:	R01 HL128370 United States HL NHLBI NIH HHS; NIH DK131177 United States DK NIDDK NIH HHS; NIH HL128370 United States HL NHLBI NIH HHS
فهرسة مساهمة:	Keywords: basement membrane; centriole; cilium; nephron; podocytes; polycystic kidney disease; renal
تواريخ الأحداث:	Date Created: 20240508 Latest Revision: 20240520
رمز التحديث:	20240520
DOI:	10.1002/cm.21870
PMID:	38715433
قاعدة البيانات:	MEDLINE

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تدمد:	1949-3592
DOI:	10.1002/cm.21870