Présentation
Lattice light-sheet microscopy1 (LLSM) is a recent fluorescence microscopy technique based on selective plane illumination microscopy2 (SPIM) that presents several key advantages: intrinsic optical sectioning, very fast imaging, low photo-toxicity and sub-micrometric spatial resolution. The great benefits of LLSM for fast 3D cellular imaging are now well established in various fields of biology.
At the Bordeaux Imaging Center (BIC) we built a home made LLSM setup that is now used in routine3. In collaboration with several research teams at the Interdisciplinary Institute for Neuroscience (IINS, UMR 5297), we perform studies in acute and organotypic rodent brain slices. Sub-micrometric neuronal compartments such as spines can be imaged down to ~ 30 µm below the tissue surface with temporal resolutions up to 200 frames/s. We demonstrate the performances of LLSM in several published and ongoing studies: measurement of AMPA receptor diffusions at spines4, vesicular transport in dendrites, spontaneous and stimulated local calcium activity in neurons and astrocytes, extracellular space and glutamate imaging5.
This presentation will first cover the principles, advantages and limitations of LLSM, and then showcase the most advanced neuroscience studies that were performed at the BIC core facility.
1. Chen BC et al. Lattice light-sheet microscopy: Imaging molecules to embryos at high spatiotemporal resolution. Science. 2014. doi:10.1126/science.1257998
2. Huisken J et al. Optical sectioning deep inside live embryos by selective plane illumination microscopy. Science. 2004. doi:10.1126/science.1100035
3. Ducros M et al. Lattice light sheet microscopy and photo-stimulation in brain slices. In: SPIE Proceedings. Vol 1086508. ; 2019:8. doi:10.1117/12.2509467
4. Getz AM et al. High-resolution imaging and manipulation of endogeneous AMPA receptor surface mobility during synaptic plasticity and learning. Sci Adv. 2022. doi:10.1126/sciadv.abm5298
5. Dembitskaya Y et al. Shadow imaging for panoptical visualization of brain tissue in vivo. Nat Commun. 2023. doi:10.1038/s41467-023-42055-2
Localisation
Bâtiment: Fernbach
Salle: Jean-Paul Aubert
Adresse: 25 Rue du Docteur Roux, Paris, France