Multiphoton microsc
opy offers unprecedented possibilities for visualizing biological processes in vivo. With its low phototoxicity and deep tissue imaging capabilities, it’s a powerful tool for elucidating the temporal dynamics of cell behaviors, thereby transforming our understanding of cardiovascular pathologies. To understand and ultimately improve cardiovascular pathologies, we aim to visualize and manipulate cells associated with the cardiovascular system in living animals using the multiphoton microscope.
In most in vivo biological studies, different animals are sacrificed at fixed time points, generating snapshots of events over time. By contrast, multiphoton microscopy allows us to appreciate the dynamics and complexity of a biological phenomenon within the same animal over time. This way, by tracking individual cells labeled with fluorescent reporters, a complete vascular pathological event can be evaluated.
High-resolution imaging permits the precise identification of any potential therapeutic mechanism and its impact on vessel functionality (hemodynamics, perfusion, and leakage). Another crucial point is cellular plasticity: various reports have identified a switch in cell lineage in pathological microenvironments, such as highly inflammatory areas. Together with genetic tools using specific promoters that direct the expression of fluorescent reporter proteins, multiphoton imaging enables us to track any desired cell type in a selected pathology and follow its behavior over time.
Confocal and Bi-photon Imaging Platform MANAGER
Leica SP8 Confocal Laser Scanning Microscope : Lasers: 405nm, Blue 488nm, Green 552nm, Red 638nm ; Detectors: PMT and HyD GaAaP
The SP8 confocal series is equipped with an upright stand, an adjustable automated stage from Scientifica, and a VIS laser set. This setup allows for the visualization of windows placed on desired animal organs without fastidious settings between experiments, enabling easy repositioning of the animal on the targeted area. The VIS single-photon laser set will be used for material settings and in specific experiments requiring high-energy photons, such as FRAP (Fluorescence Recovery After Photobleaching) or photocoagulation.
Leica SP8 Multiphoton Microscope: Equipped with a two-ray multiphoton laser beam (InSight X3, Spectra Physics), our device covers a broad excitation range in the near-infrared electromagnetic spectrum (from 680 to 1300 nm). This allows us to simultaneously excite multiple fluorophores and rapidly acquire high-resolution three-dimensional images of large tissue areas.
The DIVE detection module is the newest generation of non-descanned detectors, providing the fastest, most adaptable, and most sensitive detection system available on the market. The external (non-descanned) detectors are placed next to the objective to reduce the distance from the sample to the detector and avoid signal loss. These settings increase the sensitivity of the detectors by 2 to 3-fold compared to detectors placed in the scanner head (descanned), allowing for better sensitivity and deeper tissue penetration (up to 800µm).
Head of Platform: Yunling Xu Email: Yunling.xu@inserm.fr
Training : The platform provides comprehensive theoretical and practical training for users on the multiphoton microscope (for both slide and live imaging). We also offer continued support, accompanying users during their first few sessions until they feel confident working autonomously. Additionally, the platform offers training for procedures.
Consulting : The platform can provide assistance in developing imaging techniques and surgical procedures tailored to the specific needs of your projects. We can also provide slide acquisition, live mouse imaging, and analysis services based on your requests.
