Over the last ten years, the joint effort of the platform PLATIM and the laboratory of Plant Development and Reproduction (RDP) has developed a strong and recognized expertise in the imaging and quantification of cell mechanical properties in living plant issues. Specific pipelines have been developed to map and quantify:
1. Wall mechanical properties through direct measurements of key parameters such as wall stiffness, adhesion, and resistance to piercing, using AFMs and nano-indenters
2. Cell hydrostatic pressure through stiffness measurements made using AFMs or nano-indenters and the subsequent application of physical models but also through the development of a nano-pressure probe (in progress).
In addition of the originality of these technological approaches in mecanotransduction, the interest of our R&D teams is to develop and support their evolution directly into the environment of an IBiSA core facility. Thus, the R&D Team PLATIM/RDP is one of the rare facilities in France to provide different and integrated mechanical evaluations for both plant and animal multicellular organisms.
Publications:
- Bauer, A., Ali, O., Bied, C., Boeuf, S., Bovio, S., Delattre, A., Ingram, G., Golz, J.F. & Landrein, B. Spatiotemporally distinct responses to mechanical forces shape the developing seed of Arabidopsis. (2024) EMBO J. https://doi.org/10.1038/s44318-024-00138-w
- Creff, A., Ali, O., Bied, C., Bayle, V., Ingram, G. & Landrein, B. Evidence that endosperm turgor pressure both promotes and restricts seed growth and size. (2023) Nature Comm. https://www.nature.com/articles/s41467-022-35542-5
Lyon Multiscale Imaging Center is a microscopy facility opened to academic research and industry. It federates 3 imaging centers in the Lyon area and provides multi-scale imaging tools including light and electron microscopy:
- the PLATIM, located at the Ecole Normale Supérieure de Lyon and at Lyon-Sud Medical University, offers a wide range of light microscopy techniques ranging from fully automated video-microscopy, confocal and two-photon microscopy. The PLATIM accounts also for a platform of atomic force microscopy and microindentation for acquiring high resolution 3D topographies and for measuring micro/nano-mechanical properties.
- the Centre Technologique des Microstructures CTµ, located on the La Doua campus, is mainly an electron microscopy facility that handles multi-disciplinary projects at the interface between biology, chemistry and physics. the CTµ provides access to a wide range of scanning and transmission electron microscopy techniques, and masters cryomethods for sample preparation and observation.
- the East Lyon imaging center (CIQLE) is located in the Rockefeller Medical University. It provides access to wide-field and confocal light microscopy and transmission electron microscopy for the bio-medical research community.
Machines can be operated independently by the users after appropriate training. Users can also benefit partial or full assistance for their projects. LyMIC provides training sessions for groups or individuals.
The team is watching technology progress and implement new techniques to provide state-of-the-art tools.
The GlycoMEV team (research axis 3) aims in understanding the biosynthesis and secretion of glycoproteins with a special focus on N-glycosylation in microalgae models. As in all Eukaryotes, N-glycosylation occurs in the endoplasmic reticulum and the Golgi apparatus, where numerous enzymes act step-by-step to build and transfer oligosaccharides named N-glycans on secreted proteins. To decipher the sequence and regulation of these events, it has recently developed cryopreparation methods and imaging technologies (i.e. TEM) enabling the characterization the microalgae cell morphology and to analyse the subcellular localisation of distinct molecular actors. Furthermore, the team has acquired expertise in molecular biology techniques specific to these models allowing the generation of strains expressing various proteins fused to fluorescent reporters. They have also developed strategies based on confocal/FLIM imaging to characterize the targeting mechanisms responsible for the localization of glycoenzymes in the Golgi apparatus
SEBIO is a research unit specializing in aquatic ecotoxicology with a significant experience in the development and validation of effect-based tools for monitoring and predicting environmental impact of chemicals. As experts in marine biology, SEBIO develops in vivo whole aquatic organism imaging approaches aimed at achieving breakthrough in methods and physiological data in seawater shrimps, bivalves and fish larvae. By using light-sheet microscopy, multi-modal imaging and 3D-reconstruction, these projects will decipher fine structures and detailed homeostatic regulations, in particular related to vascular anatomy, immune cell migration and blood/hemolymph tissue perfusion, regarding infection disease, intertidal cycle, osmotic stress and blood withdrawal for laboratory sampling. In relation with INERIS, technological transfers to European environmental agencies and water monitoring programs will rely on a network of reference laboratories, research centers and organizations for environmental risk assessment.
Located in Rouen, PRIMACEN, the cell imaging facility of Normandy (1000 m2) is integrated since 2022 in HeRacLeS (Univ. Rouen Normandy, Inserm US51, CNRS UAR2026). To achieve its service offering and R&D objectives, PRIMACEN proposes a continuum with 3 pillars thanks to complementary skills in cell biology-histology, instrumentation-quality control and data processing. In tight connection with animal (rodent/marine models) and plant facilities, the first transversal pillar involves living and fixed sample preparation including labeling, for advanced light and electron microscopies. Then, appropriate resolutions are proposed to users and collaborators through multi-scale and multi-modal imaging including LSM and 2P, WF/confocal micro/macroscopy, TIRFM, FLIM-STED imaging, cryo-FLIM/confocal and TEM/cryoTEM microscopies. To finally help users in their scientific projects, image processing and analysis (3D modeling, ImageJ macros, AI…) are available on highly performing working stations.
The microscopy facility was the first technology platform established in the IBMP in 1998. Its scientific programme aims at understanding the expression of plant or animal genes over space and time at various levels. Microorganisms or biomaterials studied by partner research units are other topics of interest. Our facility follows official guidelines for « Plates-Formes Technologiques du Vivant » and has received RIO 2001, 2004 and 2006 labels. It’s part of the larger PIC.sc Strasbourg Centre Cell Imaging Facility that allows sharing devices and knowledge from several research units from CNRS, INSERM, UNISTRA. Our missions include assisting research from IBMP and partner research units, developing and implement new imaging technologies, training our user base and beyond, getting involved in microscopy education and science popularization.