Confocal light sheet microscopy: micron-scale neuroanatomy of the entire mouse brain

Silvestri, Ludovico; Bria, Alessandro; Sacconi, Leonardo; Iannello, Giulio; Pavone, Francesco S.

doi:10.1364/oe.20.020582

Cited by 212 publications

(155 citation statements)

References 49 publications

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“…The detection objective was placed on a piezo-driven mount (Physik Instrumente) to compensate for focal shift due to the refractive index mismatch between clearing solution (n BABB =1.56) and immersion medium (n air =1.00). The displacement of the objective was coupled linearly to the motion of the sample stage such that Δz PIFOC =(1-n air /n BABB ) *Δz Stage [similar to equation 1 in Silvestri et al (Silvestri et al, 2012)]. …”

Section: Imagingmentioning

confidence: 99%

Reelin and CXCL12 regulate distinct migratory behaviors during the development of the dopaminergic system

et al. 2014

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The proper functioning of the dopaminergic system requires the coordinated formation of projections extending from dopaminergic neurons in the substantia nigra (SN), ventral tegmental area (VTA) and retrorubral field to a wide array of forebrain targets including the striatum, nucleus accumbens and prefrontal cortex. The mechanisms controlling the assembly of these distinct dopaminergic cell clusters are not well understood. Here, we have investigated in detail the migratory behavior of dopaminergic neurons giving rise to either the SN or the medial VTA using genetic inducible fate mapping, ultramicroscopy, time-lapse imaging, slice culture and analysis of mouse mutants. We demonstrate that neurons destined for the SN migrate first radially and then tangentially, whereas neurons destined for the medial VTA undergo primarily radial migration. We show that tangentially migrating dopaminergic neurons express the components of the reelin signaling pathway, whereas dopaminergic neurons in their initial, radial migration phase express CXC chemokine receptor 4 (CXCR4), the receptor for the chemokine CXC motif ligand 12 (CXCL12). Perturbation of reelin signaling interferes with the speed and orientation of tangentially, but not radially, migrating dopaminergic neurons and results in severe defects in the formation of the SN. By contrast, CXCR4/CXCL12 signaling modulates the initial migration of dopaminergic neurons. With this study, we provide the first molecular and functional characterization of the distinct migratory pathways taken by dopaminergic neurons destined for SN and VTA, and uncover mechanisms that regulate different migratory behaviors of dopaminergic neurons.

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Section: Imagingmentioning

confidence: 99%

Reelin and CXCL12 regulate distinct migratory behaviors during the development of the dopaminergic system

et al. 2014

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“…69 Alternative ways to remove out-of-focus light are based on combining DSLM with structured illumination patterns 70 (see section on superresolution) or confocal slit detection. 71,72 Further improvements have been achieved with two-photon excitation 73 and/or Bessel beams. 74,75 These non-diffractive beams successfully maintain their shape and size at larger penetration depths 76 and have also been used to generate thinner light-sheets.…”

Section: B Three-dimensional Light Microscopymentioning

confidence: 99%

Invited Review Article: Advanced light microscopy for biological space research

Vos

Beghuin²,

Schwarz

et al. 2014

Review of Scientific Instruments

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As commercial space flights have become feasible and long-term extraterrestrial missions are planned, it is imperative that the impact of space travel and the space environment on human physiology be thoroughly characterized. Scrutinizing the effects of potentially detrimental factors such as ionizing radiation and microgravity at the cellular and tissue level demands adequate visualization technology. Advanced light microscopy (ALM) is the leading tool for non-destructive structural and functional investigation of static as well as dynamic biological systems. In recent years, technological developments and advances in photochemistry and genetic engineering have boosted all aspects of resolution, readout and throughput, rendering ALM ideally suited for biological space research. While various microscopy-based studies have addressed cellular response to space-related environmental stressors, biological endpoints have typically been determined only after the mission, leaving an experimental gap that is prone to bias results. An on-board, real-time microscopical monitoring device can bridge this gap. Breadboards and even fully operational microscope setups have been conceived, but they need to be rendered more compact and versatile. Most importantly, they must allow addressing the impact of gravity, or the lack thereof, on physiologically relevant biological systems in space and in ground-based simulations. In order to delineate the essential functionalities for such a system, we have reviewed the pending questions in space science, the relevant biological model systems, and the state-of-the art in ALM. Based on a rigorous trade-off, in which we recognize the relevance of multi-cellular systems and the cellular microenvironment, we propose a compact, but flexible concept for space-related cell biological research that is based on light sheet microscopy.

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“…Using wide field imaging mode, combining with tissue optical clearing techniques, imaging results of large volume sample could be acquired within a short time [6][7][8][9][10][11][12][13][14]. Minimal phototoxicity and rapid acquisition make it a powerful tool for biological studies.…”

Section: Introductionmentioning

confidence: 99%

High axial resolution imaging system for large volume tissues using combination of inclined selective plane illumination and mechanical sectioning

Zhang

Yang

et al. 2017

Biomed. Opt. Express

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Abstract:To resolve fine structures of biological systems like neurons, it is required to realize microscopic imaging with sufficient spatial resolution in three dimensional systems. With regular optical imaging systems, high lateral resolution is accessible while high axial resolution is hard to achieve in a large volume. We introduce an imaging system for high 3D resolution fluorescence imaging of large volume tissues. Selective plane illumination was adopted to provide high axial resolution. A scientific CMOS working in sub-array mode kept the imaging area in the sample surface, which restrained the adverse effect of aberrations caused by inclined illumination. Plastic embedding and precise mechanical sectioning extended the axial range and eliminated distortion during the whole imaging process. The combination of these techniques enabled 3D high resolution imaging of large tissues. Fluorescent bead imaging showed resolutions of 0.59 μm, 0.47μm, and 0.59 μm in the x, y, and z directions, respectively. Data acquired from the volume sample of brain tissue demonstrated the applicability of this imaging system. Imaging of different depths showed uniform performance where details could be recognized in either the near-soma area or terminal area, and fine structures of neurons could be seen in both the xy and xz sections.

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Confocal light sheet microscopy: micron-scale neuroanatomy of the entire mouse brain

Cited by 212 publications

References 49 publications

Reelin and CXCL12 regulate distinct migratory behaviors during the development of the dopaminergic system

Reelin and CXCL12 regulate distinct migratory behaviors during the development of the dopaminergic system

Invited Review Article: Advanced light microscopy for biological space research

High axial resolution imaging system for large volume tissues using combination of inclined selective plane illumination and mechanical sectioning

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