Setting Up a Simple Light Sheet Microscope for &lt;em&gt;In Toto&lt;/em&gt; Imaging of &lt;em&gt;C. elegans&lt;/em&gt; Development

Chardès, Claire; Mélénec, Pauline; Bertrand, Vincent; Lenne, Pierre-François

doi:10.3791/51342

Cited by 16 publications

(18 citation statements)

References 11 publications

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“…Lightsheet microscopy is also more advantageous than confocal microscopy, because it reduces photobleaching (15). Our light-sheet setup was designed from an upright microscope: a light sheet sections the sample horizontally, and the fluorescent light is collected by a high numerical aperture objective lens pointing downward (16) (Fig. 1A and Fig.…”

Section: Significancementioning

confidence: 99%

Direct laser manipulation reveals the mechanics of cell contacts in vivo

Bambardekar

Clément

Blanc

et al. 2015

Proc. Natl. Acad. Sci. U.S.A.

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Cell-generated forces produce a variety of tissue movements and tissue shape changes. The cytoskeletal elements that underlie these dynamics act at cell-cell and cell-ECM contacts to apply local forces on adhesive structures. In epithelia, force imbalance at cell contacts induces cell shape changes, such as apical constriction or polarized junction remodeling, driving tissue morphogenesis. The dynamics of these processes are well-characterized; however, the mechanical basis of cell shape changes is largely unknown because of a lack of mechanical measurements in vivo. We have developed an approach combining optical tweezers with light-sheet microscopy to probe the mechanical properties of epithelial cell junctions in the early Drosophila embryo. We show that optical trapping can efficiently deform cell-cell interfaces and measure tension at cell junctions, which is on the order of 100 pN. We show that tension at cell junctions equilibrates over a few seconds, a short timescale compared with the contractile events that drive morphogenetic movements. We also show that tension increases along cell interfaces during early tissue morphogenesis and becomes anisotropic as cells intercalate during germ-band extension. By performing pull-and-release experiments, we identify time-dependent properties of junctional mechanics consistent with a simple viscoelastic model. Integrating this constitutive law into a tissue-scale model, we predict quantitatively how local deformations propagate throughout the tissue.cell mechanics | tissue morphogenesis | optical tweezers | light-sheet microscopy | Myosin-II

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Section: Significancementioning

confidence: 99%

Direct laser manipulation reveals the mechanics of cell contacts in vivo

Bambardekar

Clément

Blanc

et al. 2015

Proc. Natl. Acad. Sci. U.S.A.

Self Cite

218

238

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“…Some of the previous research, such as OpenSPIM [39,40], had already developed stepby-step guides on how to build in-house LSFM systems. These user-friendly instructions were aimed for the spread of advanced light sheet imaging in a way of standardized design, moderate cost, and easy operation.…”

Section: D Visualization Of Branched Cells Column Using Pipmentioning

confidence: 99%

Compact plane illumination plugin device to enable light sheet fluorescence imaging of multi-cellular organisms on an inverted wide-field microscope

Guan¹,

Lee²,

Jiang³

et al. 2015

Biomed. Opt. Express

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Abstract:We developed a compact plane illumination plugin (PIP) device which enabled plane illumination and light sheet fluorescence imaging on a conventional inverted microscope. The PIP device allowed the integration of microscope with tunable laser sheet profile, fast image acquisition, and 3-D scanning. The device is both compact, measuring approximately 15 by 5 by 5 cm, and cost-effective, since we employed consumer electronics and an inexpensive device molding method. We demonstrated that PIP provided significant contrast and resolution enhancement to conventional microscopy through imaging different multi-cellular fluorescent structures, including 3-D branched cells in vitro and live zebrafish embryos. Imaging with the integration of PIP greatly reduced out-of-focus contamination and generated sharper contrast in acquired 2-D plane images when compared with the stand-alone inverted microscope. As a result, the dynamic fluid domain of the beating zebrafish heart was clearly segmented and the functional monitoring of the heart was achieved. Furthermore, the enhanced axial resolution established by thin plane illumination of PIP enabled the 3-D reconstruction of the branched cellular structures, which leads to the improvement on the functionality of the wide field microscopy. 1938-1940 (2007

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“…Hence, multi-layer mounting approaches have been developed to ensure stable conditions, such as the multi-layer approach applied by Kaufmann et al (2012) for imaging developing zebrafish embryos. LSFM has also been used to image the development of Caenorhabditis elegans embryos (Chardes et al, 2014), amoeboid movements (Takao et al, 2012), bacterial symbioses in developing zebrafish larvae (Taormina et al, 2012) and development of Tribolium castaneum embryos (Strobl and Stelzer, 2014). Despite the overwhelming focus on embryonic development, LSFM has also been used to image large cellular specimens such as three-dimensional spheroid cultures Pampaloni et al, 2013) and monitor cell division dynamics within large spheroid tumor models (Lorenzo et al, 2011).…”

Section: Sample Orientation and Preparationmentioning

confidence: 99%

Light Sheet Fluorescence Microscopy (LSFM)

et al. 2015

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The development of confocal microscopy techniques introduced the ability to optically section fluorescent samples in the axial dimension, perpendicular to the image plane. These approaches, via the placement of a pinhole in the conjugate image plane, provided superior resolution in the axial (z) dimension resulting in nearly isotropic optical sections. However, increased axial resolution, via pinhole optics, comes at the cost of both speed and excitation efficiency. Light sheet fluorescent microscopy (LSFM), a century‐old idea made possible with modern developments in both excitation and detection optics, provides sub‐cellular resolution and optical sectioning capabilities without compromising speed or excitation efficiency. Over the past decade, several variations of LSFM have been implemented each with its own benefits and deficiencies. Here we discuss LSFM fundamentals and outline the basic principles of several major light‐sheet‐based imaging modalities (SPIM, inverted SPIM, multi‐view SPIM, Bessel beam SPIM, and stimulated emission depletion SPIM) while considering their biological relevance in terms of intrusiveness, temporal resolution, and sample requirements. © 2015 by John Wiley & Sons, Inc.

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Setting Up a Simple Light Sheet Microscope for <em>In Toto</em> Imaging of <em>C. elegans</em> Development

Cited by 16 publications

References 11 publications

Direct laser manipulation reveals the mechanics of cell contacts in vivo

Direct laser manipulation reveals the mechanics of cell contacts in vivo

Compact plane illumination plugin device to enable light sheet fluorescence imaging of multi-cellular organisms on an inverted wide-field microscope

Light Sheet Fluorescence Microscopy (LSFM)

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