Live Imaging and molecular dynamics for cell, organoids, tissue and embryo : from sample preparation with access to a L2 cell culture room, to microscopy and data analysis
(Key words : spinning disk, lattice light sheet, widefield, LightSheet, FRAP, photoconversion, FCS, spot variation FCS, TIRF, single molecule, FRET, FLIM))

2 Widefield microscopes are available for our users. A Leica DMI8 Video-microscope and a Leica Thunder microscope. The Thunder is equiped with 8 Led sources of excitation:390,440,475,510,555,575,635,747 nm. Two Quad filters to spectrally separate the 8 channels on the following filters: 420-450; 462-484; 506-532; 527-551; 578-610; 602-680; 666-724; 770-850.

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 study of membrane-less compartmentalisation in cells is a growing field. At Imagerie-Gif, we have developed methods to identify these phenomena and quantify their dynamics using light microscopy techniques (DIC/FRAP/tracking, etc.).

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.

We offer workflows that enables tissular to subcellular analysis with a specialization in in immune tissues. Our team provides guidance and support at key stages throughout the process. We offer different organs fixation and sectioning methods including cryo-fixed or chemically fixed sample. Accordingly we offer various types of immunolabeling techniques using on these sections. We provide imaging services that allows to get a stitched confocal image of large samples using a high-speed confocal imaging system. Ultimately, we offer a wide range of tools enabling image restauration and analysis at the cellular and subcellular like surface quantification, intensity measurement, structures counting.
To support this workflow we rely on a fully equipped preparation laboratory and two microscopes which can be used in wide field or confocal modes using a CSU X1 Yokogawa confocal head on a Zeiss Axio Observer Z1. We have 2 computers with GPU and various analysis tools: conventional tools like Imaris, ImageJ/FIJI, Qupath, and IA tools like Ilastik, Cellpose, DeepMIB and different conda environments for Deep learning applications.