Gene expression, DNA damage, DNA structure and DNA-binding proteins structures, super-resolution (SMLM, RIM, SIM, Airyscan), single particle tracking, photo-manipulation, Correlative Light Electron Microscopy (CLEM)
Technology: Total Internal Reflection Fluorescence (TIRF)
TIRF excites fluorescence only in a very thin region near the glass surface (a few hundred nanometers). This makes it ideal for studying processes at or near the cell membrane with excellent contrast.
Numerous tests to monitor the performance of photonic microscopy systems, including light source power, observation field homogeneity, detector stability, etc. Training at PRIMACEN or at the applicants’ site.
IX83 microscope equipped with two PENTA SET TIRF, detector (sCMOS ORCA Fusion BT), TIRF module (445nm, 488nm, 514nm, 561nm, 640nm), cellVivo module, cellSens software (Evident)
Single molecule localisation microscopies in different modalities. From sample preparation, labeling, imaging to data analysis.
We use multiple imaging systems to probe the polarization of fluorescent molecules. This gives an access to their molecular organization and orientation within cells and organisms.
Resolved fluorescence acquisition combining a supercontinuum pulsed laser and wide-field detection (time-gated), with the possibility of TIRF/hilo-type optical sectioning. This device allows the observation of biosensors and/or the monitoring of interactions, and can be used in particular in the observation of the Flipper-Tr probe to provide information on local stresses.
In order to characterise dynamic phenomena at the molecular level, we have developed expertise in evaluating their diffusion (FRAP) and highlighting interactions (FRAP/FRET-FLIM).
The offer focuses on resolving molecular interactions within living cells by combining advanced quantitative microscopy methods. Techniques such as FLIM, FCS, FRAP, SPT, and super-resolution imaging enable precise analysis of interaction dynamics, diffusion, and nanoscale organization. This integrated approach provides high spatial and temporal resolution for studying biomolecular processes in their native environment.
Depending on the speed and resolution need for your live imaging project , the IGBMC platform propose different imaging systems equipped with an environmental chamber. Wide-field triggered Zeiss AxioObserver system will allow long term analysis of cell motility, wound healing application, Long-term observation of multi-labelled living cells, intracellular dynamics processes analysis (Calcium, ratiometric FRET imaging, vesicles )…High spatial 3D resolution imaging of living cells or tissues is possible on 1 point scanning (Leica TCS SP8X) or on 2 spinning disk microscopes (Yokogawa CSU-X1 or CSU-W1 scanning head) for fast acquisitions with reduced photobleaching. Live-SR modules are available on our 2-spinning disk systems enabling live Super-resolution up to 105 nm at high speed with no specific fluorophores required. 1 TIRF system equipped with a EM-CCD camera allows super resolution imaging along the optical axis (~100 nm) so imaging of membrane processes, such as receptor-ligand interactions, endocytosis, viral infection, or cell adhesion to surfaces.
2 new systems will be installed at the IGBMC platform in Spring 2025, opening up new possibilities for live imaging : 1 Lattice lightsheet will allow dynamic imaging of full sample volumes (cells, spheroids, organoids, zebrafish/c-elegans/drosophilia embryos…) with high temporal and spatial resolution and next to no phototoxicity and bleaching; 1 lattice SIM system will allow cellular, subcellular, and even sub-organelle structures in living specimens imaging in 2D and 3D with high speed, resolution below the diffraction limit (120nm, 60nm with processing) , with no need of specific fluorophores. Highest super resolution live imaging also available at our neighbored FBI Alsace Node “Plateforme Imagerie In Vitro” at INCI with the Leica Steallris STED with TauSTED and Falcon lifetime imaging option.
Several modules or lasers are available for specific photomanipulation experiments at the sub-cellular level: high energy 355 nm pulsed lasers allow uncageaging, DNA damage induction, or precise photoablation (available on a TCS SP8 confocal microscope or on fast spinning disk with Yokogawa CSU X1 scanning head), laser microdissection system, Multiphoton confocal system also available for localized damage or precise ablation, FRAP module available on 2 confocal systems and on 2 spinning disk (equipped with liveSR module), allowing the analysis of intracellular dynamics, controlled photoactivation or photoconversion applications.
Following acquisition, solution or assistance for quantitative data analysis, curve fitting and kinetics measurements can be provided.