Nanometric axial resolution of fibronectin assembly units achieved with an efficient reconstruction approach for multi-angle-TIRF microscopy

Emmanuel Soubies 1, 2 Agata Radwanska 3 Dominique Grall 3 Laure Blanc-Féraud 2 Ellen Van Obberghen-Schilling 3, 4 Sébastien Schaub 3, 2
2 MORPHEME - Morphologie et Images
CRISAM - Inria Sophia Antipolis - Méditerranée , IBV - Institut de Biologie Valrose : U1091, Laboratoire I3S - SIS - Signal, Images et Systèmes
Abstract : High resolution imaging of molecules at the cell-substrate interface is required for understanding key biological processes. Here we propose a complete pipeline for multi-angle total internal reflection fluorescence microscopy (MA-TIRF) going from instrument design and calibration procedures to numerical reconstruction. Our custom setup is endowed with a homogeneous field illumination and precise excitation beam angle. Given a set of MA-TIRF acquisitions, we deploy an efficient joint deconvolution/reconstruction algorithm based on a variational formulation of the inverse problem. This algorithm offers the possibility of using various regularizations and can run on graphics processing unit (GPU) for rapid reconstruction. Moreover, it can be easily used with other MA-TIRF devices and we provide it as an open-source software. This ensemble has enabled us to visualize and measure with unprecedented nanometric resolution, the depth of molecular components of the fibronectin assembly machinery at the basal surface of endothelial cells.
Complete list of metadatas

https://hal.inria.fr/hal-02017844
Contributor : Emmanuel Soubies <>
Submitted on : Wednesday, February 13, 2019 - 1:40:02 PM
Last modification on : Thursday, March 14, 2019 - 4:27:46 PM

Links full text

Identifiers

Collections

Citation

Emmanuel Soubies, Agata Radwanska, Dominique Grall, Laure Blanc-Féraud, Ellen Van Obberghen-Schilling, et al.. Nanometric axial resolution of fibronectin assembly units achieved with an efficient reconstruction approach for multi-angle-TIRF microscopy. Scientific Reports, Nature Publishing Group, 2019, 9 (1), ⟨10.1038/s41598-018-36119-3⟩. ⟨hal-02017844⟩

Share

Metrics

Record views

67