Optimization of 2D and 3D cell culture to study membrane organization with STED microscopy

First Authors Riccardo Maraspini
Authors Riccardo Maraspini, Chen-Ho Wang, Alf Honigmann
Corresponding Authors Alf Honigmann
Last Authors Alf Honigmann
Journal Name Journal of Physics D: Applied Physics (J Phys D: Appl Phys)
Volume 53
Issue 1
Article Number 014001
PubMed ID
WebOfScience Link WOS:000498648700001
Open Access true
Print Publication Date 2020-02-02
Online Publication Date
Abstract Epithelia cells assemble into sheets that compartmentalize organs and generate tissue barriers. This is achieved by forming polarized membrane domains, which are connected by junctional complexes. While much is known about the organization of the basal membrane due to its easy accessibility by high and super-resolution microscopy, the apical and lateral membrane domains remain poorly characterized. Here we describe our methods to study the molecular organization of apical and lateral membrane domains by combining 2D and 3D epithelial cell culture with super-resolution STED microscopy. We show that inverted cell monolayers enable live cell imaging of the apical membrane with a resolution sufficient to resolve the densely packed micro-villi of human enterocytes. Furthermore, 3D cell culture enables us to resolve adhesion complexes in the lateral domain of kidney derived cells. We envision that these methods will help to reveal the supra-molecular structure of lateral and apical membrane domains in epithelial cells.
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Affiliated With Honigmann, Predoc first author
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Publication Status Published
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DOI 10.1088/1361-6463/ab45df
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Created By thuem
Added Date 2019-12-12
Last Edited By thuem
Last Edited Date 2019-12-12 10:46:10.0
Library ID 7567
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