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Light sheet microscopy

Yicong Wu, Panagiotis Chandris, Peter W Winter, Edward Y Kim, Valentin Jaumouillé, Abhishek Kumar, Min Guo, Jacqueline M Leung, Corey Smith, Ivan Rey-Suarez, Huafeng Liu, Clare M Waterman, Kumaran S Ramamurthi, Patrick J La Riviere, Hari Shroff
Most fluorescence microscopes are inefficient, collecting only a small fraction of the emitted light at any instant. Besides wasting valuable signal, this inefficiency also reduces spatial resolution and causes imaging volumes to exhibit significant resolution anisotropy. We describe microscopic and computational techniques that address these problems by simultaneously capturing and subsequently fusing and deconvolving multiple specimen views. Unlike previous methods that serially capture multiple views, our approach improves spatial resolution without introducing any additional illumination dose or compromising temporal resolution relative to conventional imaging...
August 20, 2016: Optica
S Dini, B J Binder, S C Fischer, C Mattheyer, A Schmitz, E H K Stelzer, N G Bean, J E F Green
Automatic identification of the necrotic zone boundary is important in the assessment of treatments on in vitro tumour spheroids. This has been difficult especially when the difference in cell density between the necrotic and viable zones of a tumour spheroid is small. To help overcome this problem, we developed novel one-dimensional pair-correlation functions (PCFs) to provide quantitative estimates of the radial distance of the necrotic zone boundary from the centre of a tumour spheroid. We validate our approach on synthetic tumour spheroids in which the position of the necrotic zone boundary is known a priori It is then applied to nine real tumour spheroids imaged with light sheet-based fluorescence microscopy...
October 2016: Journal of the Royal Society, Interface
Stephanie Höhn, Armin Hallmann
BACKGROUND: The multicellular volvocine alga Pleodorina is intermediate in organismal complexity between its unicellular relative, Chlamydomonas, and its multicellular relative, Volvox, which shows complete division of labor between different cell types. The volvocine green microalgae form a group of genera closely related to the genus Volvox within the order Volvocales (Chlorophyta). Embryos of multicellular volvocine algae consist of a cellular monolayer that, depending on the species, is either bowl-shaped or comprises a sphere...
October 13, 2016: BMC Developmental Biology
Christopher Schmied, Pavel Tomancak
Light sheet fluorescent microscopy (LSFM), and in particular its most widespread flavor Selective Plane Illumination Microscopy (SPIM), promises to provide unprecedented insights into developmental dynamics of entire living systems. By combining minimal photo-damage with high imaging speed and sample mounting tailored toward the needs of the specimen, it enables in toto imaging of embryogenesis with high spatial and temporal resolution. Drosophila embryos are particularly well suited for SPIM imaging because the volume of the embryo does not change from the single cell embryo to the hatching larva...
2016: Methods in Molecular Biology
John M Heddleston, Teng-Leong Chew
Capturing dynamic processes in live samples is a nontrivial task in biological imaging. Although fluorescence provides high specificity and contrast compared to other light microscopy techniques, the photophysical principles of this method can have a harmful effect on the sample. Current advances in light sheet microscopy have created a novel imaging toolbox that allows for rapid acquisition of high-resolution fluorescent images with minimal perturbation of the processes of interest. Each unique design has its own advantages and limitations...
October 8, 2016: International Journal of Biochemistry & Cell Biology
Gagandeep Singh, Gurbir Singh, Tejwant Singh Kang
The complexation behaviour of an imidazolium based ionic liquid surfactant (ILS) 3-methyl-1-dodecylimidazolium chloride, [C12mim][Cl], and its amide and ester functionalized counterparts 3-(2-(dodecylamino)-2-oxoethyl)-1-methyl-1H-imidazol-3-ium chloride, [C12Amim][Cl], and 3-methyl-1-dodecyloxycarbonylmethylimidazolium chloride, [C12Emim][Cl], with a model protein gelatin (G) in aqueous solution has been investigated. Complexation of G with ILSs at the air-solution interface has been monitored by tensiometry, whereas complexation and ILS mediated self-assembly of G-ILS complexes in the bulk have been followed by dynamic light scattering (DLS), zeta-potential measurements, conductivity, and fluorescence techniques...
September 21, 2016: Physical Chemistry Chemical Physics: PCCP
Annette Feuchtinger, Axel Walch, Michael Dobosz
This review delves into the rapidly evolving field of deep tissue imaging at cellular resolution, reviewing popular tissue clearing and staining methods in combination with light-sheet fluorescence microscopy (LSFM) including quantification and three-dimensional visualization tools, the field of applications and perspective, particularly with the focus on preclinical cancer research and drug development. The LSFM technique presented here allows an extremely fast optical sectioning for three-dimensional reconstruction of centimeter-sized tissue samples at cellular resolution...
October 4, 2016: Histochemistry and Cell Biology
Berta Alsina, Tanya T Whitfield
The vertebrate inner ear is a precision sensory organ, acting as both a microphone to receive sound and an accelerometer to detect gravity and motion. It consists of a series of interlinked, fluid-filled chambers containing patches of sensory epithelia, each with a specialised function. The ear contains many different differentiated cell types with distinct morphologies, from the flask-shaped hair cells found in thickened sensory epithelium, to the thin squamous cells that contribute to non-sensory structures, such as the semicircular canal ducts...
September 26, 2016: Seminars in Cell & Developmental Biology
Peter H Neckel, Ulrich Mattheus, Bernhard Hirt, Lothar Just, Andreas F Mack
Novel techniques, like CLARITY and PACT, render large tissue specimens transparent and thereby suitable for microscopic analysis. We used these techniques to evaluate their potential in the intestine as an exemplary organ with a complex tissue composition. Immunohistochemistry, light sheet-, and confocal scanning-microscopy enabled us to follow complex three-dimensional structures, like nerve fibers, vessels, and epithelial barriers throughout the entire organ. Moreover, in a systematic electron microscopic study, we analyzed the morphology and preservation of tissue on ultrastructural level during the clearing process...
September 29, 2016: Scientific Reports
Abhishek Kumar, Ryan Christensen, Min Guo, Panos Chandris, William Duncan, Yicong Wu, Anthony Santella, Mark Moyle, Peter W Winter, Daniel Colón-Ramos, Zhirong Bao, Hari Shroff
Dual-view inverted selective plane illumination microscopy (diSPIM) enables high-speed, long-term, four-dimensional (4D) imaging with isotropic spatial resolution. It is also compatible with conventional sample mounting on glass coverslips. However, broadening of the light sheet at distances far from the beam waist and sample-induced scattering degrades diSPIM contrast and optical sectioning. We describe two simple improvements that address both issues and entail no additional hardware modifications to the base diSPIM...
August 2016: Biological Bulletin
Stephan Daetwyler, Jan Huisken
In light sheet microscopy, optical sectioning by selective fluorescence excitation with a sheet of light is combined with fast full-frame acquisition. This illumination scheme provides minimal photobleaching and phototoxicity. Complemented with remote focusing and multi-view acquisition, light sheet microscopy is the method of choice for acquisition of very fast biological processes, large samples, and high-throughput applications in areas such as neuroscience, plant biology, and developmental biology. This review explains why light sheet microscopes are much faster and gentler than other established fluorescence microscopy techniques...
August 2016: Biological Bulletin
Sawetree Pakkarato, Wipawee Thoungseabyoun, Apussara Tachow, Atsara Rawangwong, Yoshiteru Kagawa, Yuji Owada, Hisatake Kondo, Wiphawi Hipkaeo
In our previous immuno-light microscopic study with an antibody for fatty acid binding protein of type 7 or brain type (FABP-7, B-FABP), the adrenomedullary sustentacular cells were revealed to have secondary processes that present faint immunostaining and an ill-defined sheet-like appearance, in addition to the well-recognized primary processes that present distinct immunostaining and a fibrous appearance. The secondary processes were regarded as corresponding to known ultrastructural profiles of sustentacular cells with a thickness of less than 0...
September 15, 2016: Anatomical Science International
James D Manton, Eric J Rees
We propose a three-objective light sheet microscopy geometry which, through a combination of skewed lattice light sheet excitation through two objectives and the computational fusion of images taken from two separate lens pairings, would allow for isotropic super-resolution in mesoscopic samples. We also show that simultaneous coherent excitation through two excitation objectives could further substantially increase resolution. Simulations demonstrate that our design could achieve a resolution of 120 nm for EGFP imaging while minimizing photodamage...
September 15, 2016: Optics Letters
Patrick Theer, Denitsa Dragneva, Michael Knop
Light-sheet fluorescence microscopy (LSFM), also termed single plane illumination microscopy (SPIM), enables live cell fluorescence imaging with optical sectioning capabilities superior to confocal microscopy and without any out-of-focus exposure of the specimen. However, the need of two objective lenses, one for light-sheet illumination and one for imaging, imposes geometrical constraints that require LSFM setups to be adapted to the specific needs of different types of specimen in order to obtain optimal imaging conditions...
2016: Scientific Reports
Devynn M Wulstein, Kathryn E Regan, Rae M Robertson-Anderson, Ryan McGorty
Using light-sheet microscopy combined with digital Fourier methods we probe the dynamics of colloidal samples and DNA molecules. This combination, referred to as selective-plane illumination differential dynamic microscopy (SPIDDM), has the benefit of optical sectioning to study, with minimal photobleaching, thick samples allowing us to measure the diffusivity of colloidal particles at high volume fractions. Further, SPIDDM exploits the inherent spatially-varying thickness of Gaussian light-sheets. Where the excitation sheet is most focused, we capture high spatial frequency dynamics as the signal-to-background is high...
September 5, 2016: Optics Express
Markus Aswendt, Martin Schwarz, Walid M Abdelmoula, Jouke Dijkstra, Stefanie Dedeurwaerdere
Magnetic resonance imaging, positron emission tomography, and optical imaging have emerged as key tools to understand brain function and neurological disorders in preclinical mouse models. They offer the unique advantage of monitoring individual structural and functional changes over time. What remained unsolved until recently was to generate whole-brain microscopy data which can be correlated to the 3D in vivo neuroimaging data. Conventional histological sections are inappropriate especially for neuronal tracing or the unbiased screening for molecular targets through the whole brain...
September 2, 2016: Molecular Imaging and Biology: MIB: the Official Publication of the Academy of Molecular Imaging
Filippo Piccinini, Anna Tesei, Alessandro Bevilacqua
BACKGROUND: Cancer multicellular spheroids are commonly used as 3D tumour models for testing drugs and radiotherapy treatments. The volume plays a key role in analysis of the results. Several methods have been proposed in the literature to compute the spheroid's volume from one 2D microscopy image (i.e. a single projection). However, the literature lacks reviews summarising the different methods available. Furthermore, there are no well-established approaches by which to compare the different methods and determine the best one...
October 2016: Computer Methods and Programs in Biomedicine
Deirdre Healy, Maria E Nash, Alexander Gorelov, Kerry Thompson, Peter Dockery, Sergey Beloshapkin, Yury Rochev
This study describes the development and cell culture application of nanometer thick photocrosslinkable thermoresponsive polymer films prepared by physical adsorption. Two thermoresponsive polymers, poly(N-isopropylacrylamide (NIPAm)-co-acrylamidebenzophenone (AcBzPh)) and poly(NIPAm-co-AcBzPh-co-N-tertbutylacrylamide) are investigated. Films are prepared both above and below the polymers' lower critical solution temperatures (LCSTs) and cross-linked, to determine the effect, adsorption preparation temperature has on the resultant film...
August 31, 2016: Macromolecular Bioscience
Nelson Monteiro, Elizabeth E Smith, Shantel Angstadt, Weibo Zhang, Ali Khademhosseini, Pamela C Yelick
Tissue engineering and regenerative medicine technologies offer promising therapies for both medicine and dentistry. Our long-term goal is to create functional biomimetic tooth buds for eventual tooth replacement in humans. Here, our objective was to create a biomimetic 3D tooth bud model consisting of dental epithelial (DE) - dental mesenchymal (DM) cell sheets (CSs) combined with biomimetic enamel organ and pulp organ layers created using GelMA hydrogels. Pig DE or DM cells seeded on temperature-responsive plates at various cell densities (0...
November 2016: Biomaterials
Wiebke Jahr, Benjamin Schmid, Michael Weber, Jan Huisken
LIGHT SHEET MICROSCOPY IN THE MUSEUM: Light sheet microscopy (or selective plane illumination microscopy) is an important imaging technique in the life sciences. At the same time, this technique is also ideally suited for community outreach projects, because it produces visually appealing, highly dynamic images of living organisms and its working principle can be understood with basic optics knowledge. Still, the underlying concepts are widely unknown to the non-scientific public. On the occasion of the UNESCO International Year of Light, a technical museum in Dresden, Germany, launched a special, interactive exhibition...
2016: PloS One
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