Atomic force microscopy on living cells or 3D samples is a powerful technique to measure the mechanical properties such as elastic moduli , rigidity, adhesion force, adhesion energy. Combining optical images, AI and AFM, cell population’s mechanome becomes accessible. 3D samples (tissues, organoids) mechanical properties require specific expertise we also provide.
Technology: Brightfield microscopy
Brightfield microscopy is the most widely used optical microscopy technique. A beam of white light passes through the sample, and the contrast in the image comes from the way different parts of the specimen absorb or scatter this light. This straightforward method makes it possible to observe the general structure, shape, and organization of cells and tissues.
Inflammatory and cardiovascular diseases are complex, multifactorial conditions that affect multiple organs and cellular components. To tackle these intricate biological challenges, we employ and advance multi-modal and multi-scale imaging and flow cytometry pipelines. These pipelines include the entire process from sample preparation, image acquisition to image analysis and quantification. Our workflows are adpated to various tissues, including the heart, kidneys, blood vessels, muscles, liver and adipose tissue. They operate across scales ranging from whole organs to subcellular levels, employing techniques such as widefield imaging, confocal spectral imaging, high-resolution imaging (Airyscan), high-content imaging (Axioscan 7, Opera Phenix), and cytometry-based analysis (spectral, sorting, and analysis).
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.
Workflow from sample preparation, MET and cryo-confocal, advanced photonic microscopy
WF/confocal/FLIM, CLEM, image analysis (AMIRA)
Zeiss CD7, XY, Z-stacks, multi-points, mosaics, time-lapse
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))
Histology : from sample preparation with the RHEM and PHIV facility (human, animal and plant tissue) to digitisation of virtual slides and image analysis
(Key words : brightfield and multiplex staining, Nanozoomer (brightfield), Axioscan (fluorescence), Pannoramic (brightfield and fluorescence), QuPath and Histometrix with machine/deep learning)
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 imaging system Kratoscope acquires an image of the organ in the block of tissue each time a slice is cut by a microtome (FFPE blocks) or a cryostat (frozen tissue), in a fully automated process. By getting a stack of slices at increasing depth, it is then possible to reconstruct a 3D view of the organ.
In addition to a possible fluorescent signal detection, the contrast of the images comes from the difference in autofluorescence of different structures of the organ and highlights anatomical its anatomy. It can provide information not visible on micro CT images, when small structures lack radio contrast.
support from sample preparation to image analysis – including visualisation of organoids/spheroids in 3D imaging. Pool of systems and expertise for these studies (transparization, staining, light sheets, etc.).