VICENTE, Filipe Nunes; MASSOU, Sophie; WETZEL, Franziska; MEHIDI, Amine; STREHLE, Dan; LEDUC, Cecile; VOITURIEZ, Raphaël; ROSSIER, Olivier; NASSOY, Pierre; GIANNONE, Grégory

doi:10.21203/rs.3.pex-961/v1

Métadonnées

Afficher la notice complète

Licence d’utilisation du document

VICENTE, Filipe Nunes
Interdisciplinary Institute for Neuroscience [Bordeaux] [IINS]

MASSOU, Sophie
Interdisciplinary Institute for Neuroscience [Bordeaux] [IINS]

WETZEL, Franziska
Laboratoire Photonique, Numérique et Nanosciences [LP2N]

Langue

Document de travail - Pré-publication

Résumé en anglais

Cell mechano-sensing is based on biomolecule deformations and reorganizations, yet the molecular mechanisms are still unclear. Super-resolution microscopy (SRM) and single protein tracking (SPT) techniques reveal the dynamic organization of proteins at the nanoscale. In parallel, stretchable substrates are used to investigate cellular responses to mechanical forces. However, simultaneous combination of SRM/SPT and cell stretching has never been achieved. Here, we present a cell stretching device compatible with SRM and SPT, composed of an ultra-thin Polydimethylsiloxane (PDMS) layer. The PDMS sheet is gliding on a glycerol-lubricated glass cover-slip to ensure flatness during uniaxial stretching, generated with a 3D-printed micromechanical device by a mobile arm connected to a piezoelectric translator. This method enables to obtain super-resolved images of protein reorganization after live stretching, and to monitor single protein deformation and recruitment inside mechanosensitive structures upon stretching. This protocol is related to the publication ‘Cell stretching is amplified by active actin remodeling to deform and recruit proteins in mechanosensitive structures’, in Nature Cell Biology.< Réduire

Mots clés en anglais

cytoskeleton

cell stretching

superresolution microscopy

single particle tracking

mechanobiology

mechanosensing

focal adhesions

Métadonnées

Partager cette publication !

Licence d’utilisation du document

A micromechanical cell stretching device compatible with super-resolution microscopy and single protein tracking

Langue

Résumé en anglais

Mots clés en anglais

URI

DOI

Origine

Unités de recherche