A5057529735- Advanced Fluorescence Microscopy Techniques
- Cell Image Analysis Techniques
- Photoacoustic and Ultrasonic Imaging
- Optical Coherence Tomography Applications
- Neural dynamics and brain function
- Neuroscience and Neural Engineering
- Photoreceptor and optogenetics research
- Spectroscopy Techniques in Biomedical and Chemical Research
- Nonlinear Optical Materials Studies
- Advanced Optical Sensing Technologies
- Advanced Electron Microscopy Techniques and Applications
- Advanced Biosensing Techniques and Applications
- Nanofabrication and Lithography Techniques
- Digital Holography and Microscopy
- Optical Imaging and Spectroscopy Techniques
- Single-cell and spatial transcriptomics
- Angiogenesis and VEGF in Cancer
- Intraocular Surgery and Lenses
- Laser Material Processing Techniques
- Visual perception and processing mechanisms
- Neuroscience and Neuropharmacology Research
- Photodynamic Therapy Research Studies
- Neurogenesis and neuroplasticity mechanisms
- Retinal Diseases and Treatments
- Advanced Memory and Neural Computing
University of California, Berkeley
2021-2023
Inserm
2016-2021
École Polytechnique
2016-2021
Centre National de la Recherche Scientifique
2016-2021
Université Paris-Saclay
2017-2020
Institut de la Vision
2016
Centrale Marseille
2015
Institut Fresnel
2015
Aix-Marseille Université
2015
Abstract Astrocytes play essential roles in the neural tissue where they form a continuous network, while displaying important local heterogeneity. Here, we performed multiclonal lineage tracing using combinatorial genetic markers together with new large volume color imaging approach to study astrocyte development mouse cortex. We show that cortical clones intermix their neighbors and display extensive variability terms of spatial organization, number subtypes cells generated. Clones develop...
Abstract Two-photon imaging of endogenous fluorescence can provide physiological and metabolic information from intact tissues. However, simultaneous multiple intrinsic fluorophores, such as nicotinamide adenine dinucleotide(phosphate) (NAD(P)H), flavin dinucleotide (FAD) retinoids in living systems is generally hampered by sequential multi-wavelength excitation resulting motion artifacts. Here, we report on efficient multicolor two-photon fluorophores with absorption spectra spanning the...
Multiphoton microscopy combined with genetically encoded fluorescent indicators is a central tool in biology. Three-photon (3P) excitation the short-wavelength infrared (SWIR) water transparency bands at 1.3 and 1.7 µm opens up new opportunities for deep-tissue imaging. However, novel strategies are needed to enable in-depth multicolor fluorescence imaging fully develop such an approach. Here, we report on multiband SWIR source that simultaneously emits ultrashort pulses has characteristics...
Single-cell dry mass measurement is used in biology to follow cell cycle, address effects of drugs, or investigate metabolism. Quantitative phase imaging technique with quadriwave lateral shearing interferometry (QWLSI) allows measuring mass. The very simple set up, as it integrated a camera-like instrument. It simply plugs onto standard microscope and uses white light illumination source. Its working principle first explained, from image acquisition automated segmentation algorithm...
Abstract Large-scale microscopy approaches are transforming brain imaging, but currently lack efficient multicolor contrast modalities. We introduce chromatic multiphoton serial (ChroMS) microscopy, a method integrating one‐shot excitation through wavelength mixing and block-face image acquisition. This approach provides organ-scale micrometric imaging of spectrally distinct fluorescent proteins label-free nonlinear signals with constant micrometer-scale resolution sub-micron channel...
The biophysical properties of existing optogenetic tools constrain the scale, speed, and fidelity precise control. Here, we use structure-guided mutagenesis to engineer opsins that exhibit very high potency while retaining fast kinetics. These new enable large-scale, temporally spatially control population neural activity. We extensively benchmark these against provide a detailed characterization diverse family under two-photon illumination. This establishes resource for matching optimal...
The spectral window lying between 1.6 and 1.7 μm is interesting for in-depth multiphoton microscopy of intact tissues due to reduced scattering absorption in this wavelength range. However, wide adoption excitation range will rely on the availability robust cost-effective high peak power pulsed lasers operating at these wavelengths. In communication, we report a monolithically integrated repetition rate (50 MHz) all-fiber femtosecond laser based soliton self-frequency shift providing 9 nJ,...
Improving the imaging speed of multiphoton microscopy is an active research field. Among recent strategies, light-sheet illumination holds distinctive advantages for achieving fast in vivo . However, photoperturbation remains poorly investigated. We show here that heart beat rate zebrafish embryos a sensitive probe linear and nonlinear photoperturbations. By analyzing its behavior with respect to laser power, pulse frequency wavelength, we derive guidelines find best balance between signal...
Summary Patterned optogenetic activation of defined neuronal populations in the intact brain can reveal fundamental aspects neural codes perception and behavior. The biophysical properties existing tools, however, constrain scale, speed, fidelity precise optical control. Here we use structure-guided mutagenesis to engineer opsins that exhibit very high potency while retaining fast kinetics. These new enable large-scale, temporally spatially control population activity vivo vitro . We...
Brain computation depends on intricately connected yet highly distributed neural networks. Due to the absence of requisite technologies, causally testing fundamental hypotheses nature inter-areal processing have remained largely out-of-each. Here we developed first two photon holographic mesoscope, a system capable simultaneously reading and writing activity patterns with single cell resolution across large regions brain. We demonstrate precise photo-activation spatial temporal sequences...
Abstract When sensory information is incomplete or ambiguous, the brain relies on prior expectations to infer perceptual objects. Despite centrality of this process perception, neural mechanism inference not known. Illusory contours (ICs) are key tools study because they contain edges objects that implied only by their spatial context. Using cellular resolution, mesoscale two-photon calcium imaging and multi-Neuropixels recordings in mouse visual cortex, we identified a sparse subset neurons...
Get PDF Email Share with Facebook Tweet This Post on reddit LinkedIn Add to CiteULike Mendeley BibSonomy Citation Copy Text C. Stringari, L. Abdeladim, P. Mahou, G. Malkinson, S. Brizion, J. Galey, W. Supatto, R. Legouis, A. Pena, and E. Beaurepaire, "Multicolor two-photon imaging of endogenous fluorophores in living tissues by wavelength mixing," Biophotonics Congress: Biomedical Optics Congress 2018 (Microscopy/Translational/Brain/OTS), OSA Technical Digest (Optica Publishing Group, 2018),...
The cells composing brain tissue, neurons, and glia, form extraordinarily complex networks that support cognitive functions. Understanding the organization development of these requires quantitative data resolved at single cell level. To this aim, we apply novel large-scale 3D multicolor microscopy methodologies in combination with "Brainbow", a transgenic approach enabling to label neural diverse combinations spectrally distinct fluorescent proteins. In paper, present pipeline based on...
Protoplasmic astrocytes (PrA) located in the mouse cerebral cortex are tightly juxtaposed, forming an apparently continuous three-dimensional matrix at adult stages. Thus far, no immunostaining strategy can single them out and segment their morphology mature animals over course of corticogenesis. Cortical PrA originate from progenitors dorsal pallium easily be targeted using utero electroporation integrative vectors. A protocol is presented here to label these cells with multiaddressable...
We present the first two-photon holographic mesoscope, a system allowing to selectively photostimulate ensembles of neurons with high spatiotemporal resolution while recording from thousands across several surrounding areas.
One optimal window for biological imaging lies around 1675 ± 20 nm wavelength, a result of the decreased effects both tissue scattering and water absorption. Furthermore, in nonlinear microscopy signal-to-background ratio, which limits maximum penetration depth, is dramatically increased with increasing photon order. As consequence, three-photon an excitation wavelength approximately was recently proposed as promising approach deep microscopic live tissues. Wide adoption this technique will...
Protoplasmic astrocytes (PrA) located in the mouse cerebral cortex are tightly juxtaposed, forming an apparently continuous three-dimensional matrix at adult stages. Thus far, no immunostaining strategy can single them out and segment their morphology mature animals over course of corticogenesis. Cortical PrA originate from progenitors dorsal pallium easily be targeted using utero electroporation integrative vectors. A protocol is presented here to label these cells with multiaddressable...
Light-sheet fluorescence microscopy is a method of choice for multiscale live imaging. Indeed, its orthogonal geometry results in high acquisition speed, large field-of-view and low photodamage. Its combination with multiphoton excited improves imaging depth biological tissues. However, it appears femtosecond laser sources commonly used at an 80 MHz repetition rate may not be optimized to take full advantage light-sheet illumination during Hence, we investigated the nature induced...