Microscopic Imaging
Our laboratory has unique equipment and expertise for the study of diseases of the nervous system using advanced imaging.
Team
Operational Manager. Clémence Mille (Assistante Ingénieure, Clemence.Mille@inserm.fr)
Scientific manager. Christian Specht (CRCN, Christian.Specht@inserm.fr)
Confocal Microscopy managers. Julien Giustiniani ( Institut Baulieu, Julien.Giustiniani@inserm.fr) – Marcel Tawk (CRCN, Marcel.Tawk@inserm.fr)
Super-resolution Microscopy
This platform provides super-resolution microscopy equipment (SMLM, SIM) which makes it possible to study neuronal structures with a spatial resolution that exceeds the diffraction limits. Among these technologies, single molecule localization microscopy (SMLM) achieves a precision of the order of 10 nanometers, which allows ultra-structural characterization of neurons as well as quantification by molecular counting and spatio-temporal tracking of targets. specific in living cells.
Activities
This platform aims to study pathological processes in neurons using super-resolution imaging approaches (SIM, SMLM).
SIM microscopy (structured illumination microscopy) is based on varied illumination of the field of view which provides access to information in frequency space by Fourier transformation, thus improving the resolution to around 100 nm. This allows the characterization of fine structural details, as illustrated by the identification of spectrin rings in the initial segment of the axon.
SMLM (single molecule localization microscopy) is based on the principle of detecting single molecules, which can be located with a spatial precision of around ten nanometers. Among the different approaches, dSTORM generally consists of immunostaining of endogenous targets using organic dyes. In the PALM configuration, photo-convertible fluorescent proteins are used to label and visualize targets in living cells.
Financements
Equipements
Our laboratory is equipped with a ZEISS Elyra PS1 microscopy station which allows two types of super-resolution imaging, SIM and SMLM, including PALM and dSTORM. Moreover, an Abbelight SAFe nanoscope offers additional SMLM options, including spectral unmixing and astigmatic imaging for three-dimensional localization of single molecule detections.
Access
This equipment is available to external users on request, generally within the framework of scientific collaboration. Submitted projects are evaluated by a scientific commission made up of experts in different imaging technologies. Contact: christian.specht@inserm.fr
Microscopie CLEM (Correlative Light and Electron Microscopy)
This platform managed by the CryoCapCell Company (Mr. Belle, X Heiligenstein) offers methodologies, equipment and expertise to carry out multimodal imaging analyzes, with a particular accent on Clem correlative microscopy, which combines photonic and electronic microscopies.
Activities
Correlative optical and electron microscopy makes it possible to simultaneously obtain information from optical microscopy (localization of individual proteins in their hydrated native environment) and electron microscopy (near-atomic resolution). The technology developed makes it possible to combine a fluorescence microscope with a Live µ High Pressure Freezer which makes it possible to obtain a time correlation of <2s between live imaging and the vitrification of a thick sample (200µm) by freezing at high pressure.
Confocal microscopy
Our laboratory has two confocal microscopes.
A Leica SP8 confocal microscope, equipped with modules for spatio-temporal control of illumination, thus enabling opto-genetic approaches such as decapping and dynamic measurements of protein populations (FRAP) in living cells.
A confocal spinning disk microscope (managed by the UMS44) allows you to observe living samples (cells, organoids, small animals). The “spinning-disk” uses lasers to illuminate samples over the entire observation area simultaneously. It reduces acquisition time and photobleaching of fluorescent samples as well as phototoxicity resulting from illumination.
Electron microscopy
Electron microscopy is an essential technology in neuroscience research. Its nanometric resolution allows a detailed study of the structure and constituents of neurons, glial cells and myelin sheaths. Our unit has a 100 kV transmission electron microscope (JEM 1010, JEOL) equipped with a digital camera (Erlangshen ES500W), intensively used by researchers and clinicians.
Optical microscopy
Our laboratory has the latest generation optical microscopes for histological analysis.