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Accueil du site > Animations Scientifiques > Séminaires 2013 > Jean-Baptiste Sibarita - Resolving molecular organization and dynamics of post-synaptic molecules using localization-based super-resolution microscopy

Jean-Baptiste Sibarita - Resolving molecular organization and dynamics of post-synaptic molecules using localization-based super-resolution microscopy

Speaker :Jean-Baptiste Sibarita - UMR5297-CNRS, Univ. Bordeaux, Interdisciplinary Institute for NeuroScience, France

When : thursday 30 th May at 11 am

Where : Salle CO23 (grande salle de réunions du CBP rez-de-chaussée LR6)

Title :Resolving molecular organization and dynamics of post-synaptic molecules using localization-based super-resolution microscopy

In recent years, super-resolution optical microscopy techniques have been developed and proven to break the resolution limit given by the diffraction of light in optical systems (typically 250 nm). Among these, those based on the sequential stochastic photo-conversion of sparse subsets of single fluorophores, i.e. (f)PALM, (d)STORM or GSDIM, rely on the ability of determining the center of the point spread function (PSF) created by each single point emitter. These techniques have become extremely popular due to their affordability and relatively simple implementation on a conventional microscope. They allow the localization and tracking of a large number of biomolecules with close to molecular accuracy (down to 10 nm in lateral and 40 nm in axial) and millisecond scale temporal resolution. Nevertheless, a major step relies in the image analysis, often time-consuming and not easy to handle by non-specialists.

We will first present some practical implementations enabling the use of such powerful techniques in routine. Then, we will present a biological application to monitor the organization and dynamics of post-synaptic receptors within live neurons. Using independent super-resolution light imaging methods and electron microscopy, we will show that AMPA receptors are highly concentrated in nanodomains, instead of diffusively distributed in the PSD as generally thought, leading to important consequences on our understanding of excitatory neurotransmission.

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