Cargando…

Combining patch-clamping and fluorescence microscopy for quantitative reconstitution of cellular membrane processes with Giant Suspended Bilayers

In vitro reconstitution and microscopic visualization of membrane processes is an indispensable source of information about a cellular function. Here we describe a conceptionally novel free-standing membrane template that facilitates such quantitative reconstitution of membrane remodelling at differ...

Descripción completa

Detalles Bibliográficos
Autores principales: Velasco-Olmo, Ariana, Ormaetxea Gisasola, Julene, Martinez Galvez, Juan Manuel, Vera Lillo, Javier, Shnyrova, Anna V.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Nature Publishing Group UK 2019
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6510758/
https://www.ncbi.nlm.nih.gov/pubmed/31076583
http://dx.doi.org/10.1038/s41598-019-43561-4
_version_ 1783417460900233216
author Velasco-Olmo, Ariana
Ormaetxea Gisasola, Julene
Martinez Galvez, Juan Manuel
Vera Lillo, Javier
Shnyrova, Anna V.
author_facet Velasco-Olmo, Ariana
Ormaetxea Gisasola, Julene
Martinez Galvez, Juan Manuel
Vera Lillo, Javier
Shnyrova, Anna V.
author_sort Velasco-Olmo, Ariana
collection PubMed
description In vitro reconstitution and microscopic visualization of membrane processes is an indispensable source of information about a cellular function. Here we describe a conceptionally novel free-standing membrane template that facilitates such quantitative reconstitution of membrane remodelling at different scales. The Giant Suspended Bilayers (GSBs) spontaneously swell from lipid lamella reservoir deposited on microspheres. GSBs attached to the reservoir can be prepared from virtually any lipid composition following a fast procedure. Giant unilamellar vesicles can be further obtained by GSB detachment from the microspheres. The reservoir stabilizes GSB during deformations, mechanical micromanipulations, and fluorescence microscopy observations, while GSB-reservoir boundary enables the exchange of small solutes with GSB interior. These unique properties allow studying macro- and nano-scale membrane deformations, adding membrane-active compounds to both sides of GSB membrane and applying patch-clamp based approaches, thus making GSB a versatile tool for reconstitution and quantification of cellular membrane trafficking events.
format Online
Article
Text
id pubmed-6510758
institution National Center for Biotechnology Information
language English
publishDate 2019
publisher Nature Publishing Group UK
record_format MEDLINE/PubMed
spelling pubmed-65107582019-05-23 Combining patch-clamping and fluorescence microscopy for quantitative reconstitution of cellular membrane processes with Giant Suspended Bilayers Velasco-Olmo, Ariana Ormaetxea Gisasola, Julene Martinez Galvez, Juan Manuel Vera Lillo, Javier Shnyrova, Anna V. Sci Rep Article In vitro reconstitution and microscopic visualization of membrane processes is an indispensable source of information about a cellular function. Here we describe a conceptionally novel free-standing membrane template that facilitates such quantitative reconstitution of membrane remodelling at different scales. The Giant Suspended Bilayers (GSBs) spontaneously swell from lipid lamella reservoir deposited on microspheres. GSBs attached to the reservoir can be prepared from virtually any lipid composition following a fast procedure. Giant unilamellar vesicles can be further obtained by GSB detachment from the microspheres. The reservoir stabilizes GSB during deformations, mechanical micromanipulations, and fluorescence microscopy observations, while GSB-reservoir boundary enables the exchange of small solutes with GSB interior. These unique properties allow studying macro- and nano-scale membrane deformations, adding membrane-active compounds to both sides of GSB membrane and applying patch-clamp based approaches, thus making GSB a versatile tool for reconstitution and quantification of cellular membrane trafficking events. Nature Publishing Group UK 2019-05-10 /pmc/articles/PMC6510758/ /pubmed/31076583 http://dx.doi.org/10.1038/s41598-019-43561-4 Text en © The Author(s) 2019 Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.
spellingShingle Article
Velasco-Olmo, Ariana
Ormaetxea Gisasola, Julene
Martinez Galvez, Juan Manuel
Vera Lillo, Javier
Shnyrova, Anna V.
Combining patch-clamping and fluorescence microscopy for quantitative reconstitution of cellular membrane processes with Giant Suspended Bilayers
title Combining patch-clamping and fluorescence microscopy for quantitative reconstitution of cellular membrane processes with Giant Suspended Bilayers
title_full Combining patch-clamping and fluorescence microscopy for quantitative reconstitution of cellular membrane processes with Giant Suspended Bilayers
title_fullStr Combining patch-clamping and fluorescence microscopy for quantitative reconstitution of cellular membrane processes with Giant Suspended Bilayers
title_full_unstemmed Combining patch-clamping and fluorescence microscopy for quantitative reconstitution of cellular membrane processes with Giant Suspended Bilayers
title_short Combining patch-clamping and fluorescence microscopy for quantitative reconstitution of cellular membrane processes with Giant Suspended Bilayers
title_sort combining patch-clamping and fluorescence microscopy for quantitative reconstitution of cellular membrane processes with giant suspended bilayers
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6510758/
https://www.ncbi.nlm.nih.gov/pubmed/31076583
http://dx.doi.org/10.1038/s41598-019-43561-4
work_keys_str_mv AT velascoolmoariana combiningpatchclampingandfluorescencemicroscopyforquantitativereconstitutionofcellularmembraneprocesseswithgiantsuspendedbilayers
AT ormaetxeagisasolajulene combiningpatchclampingandfluorescencemicroscopyforquantitativereconstitutionofcellularmembraneprocesseswithgiantsuspendedbilayers
AT martinezgalvezjuanmanuel combiningpatchclampingandfluorescencemicroscopyforquantitativereconstitutionofcellularmembraneprocesseswithgiantsuspendedbilayers
AT veralillojavier combiningpatchclampingandfluorescencemicroscopyforquantitativereconstitutionofcellularmembraneprocesseswithgiantsuspendedbilayers
AT shnyrovaannav combiningpatchclampingandfluorescencemicroscopyforquantitativereconstitutionofcellularmembraneprocesseswithgiantsuspendedbilayers