Time and polarisation resolved microscopy to follow proteins interactions in neurobiology
type: Dataset
description: In the framework of this thesis, we have used FRET (Forster Resonance Energy Transfer) as a mechanism to follow the interaction of proteins from the plasma membrane to the cytoplasm of cells. To quantify FRET, we have chosen Fluorescence Lifetime Imaging Microscopy (FLIM) since this method is independent of the concentration and intensity of the fluorophores. To have a good axial resolution, a TIRFLIM set-up (Total Internal Reflection Fluorescence Lifetime Imaging Microscopy) was developed and this allowed us to perform wide-field imaging with sub-wavelength axial resolution. This set-up was calibrated and optimized in order to answer biological questions. Different approaches were tested in order to measure the penetration depth of the evanescent field and especially plasmonic surfaces were used to further enhance the axial resolution. Our set-up was dedicated to the study of the effect of cholesterol on the interaction between the Amyloid Precursor Protein (APP), a transmembrane protein involved in Alzheimer Disease, and one of its cleaving enzyme (BACE1). We performed a dynamic tracking of APP and BACE1 proximity under the effect of cholesterol, in HEK-293 cells and primary cultures of embryonic rat hippocampal neurons, thanks to our TIRFLIM set-up.Time-resolved fluorescence anisotropy has been implemented on our set-up. This has enabled us to measure the rotational correlation time of fluorophores and to investigate quantitatively different states of homodimerization of proteins involved in Alzheimer’s disease.
identifier: NNT: 2011PA112315
issued: 2026-05-09T20:09:04.339861
language: fr
modified: 2026-05-09T20:09:04.339866
contact point: Organization
  fn: CCSD
keyword: anisotropie-de-fluorescence
keyword: flim
keyword: fluorescence-anisotropy
keyword: fret
keyword: homofret
keyword: infoeu-reposemanticsdoctoralthesis
keyword: interactions-proteiques
keyword: neurobiologie
keyword: neurobiology
keyword: physcondcm-genphysics-physicscondensed-matter-cond-matother-cond-matother
keyword: plasmonic-surfaces
keyword: proteins-interactions
keyword: surfaces-plasmoniques
keyword: theses
keyword: tirf
landing page: https://pastel.hal.science/tel-00859360

Time and polarisation resolved microscopy to follow proteins interactions in neurobiology
type: Distribution
format: HTML
issued: 2026-05-09T20:09:04.343056
modified: 2026-05-09T20:09:04.326710
access URL: https://pastel.hal.science/tel-00859360

test_moissonnage_selune
type: Agent