The Experts below are selected from a list of 47706 Experts worldwide ranked by ideXlab platform
David W Mccurdy - One of the best experts on this subject based on the ideXlab platform.
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high resolution Confocal Imaging of wall ingrowth deposition in plant transfer cells semi quantitative analysis of phloem parenchyma transfer cell development in leaf minor veins of arabidopsis
BMC Plant Biology, 2015Co-Authors: Suong T T Nguyen, David W MccurdyAbstract:Transfer cells (TCs) are trans-differentiated versions of existing cell types designed to facilitate enhanced membrane transport of nutrients at symplasmic/apoplasmic interfaces. This transport capacity is conferred by intricate wall ingrowths deposited secondarily on the inner face of the primary cell wall, hence promoting the potential trans-membrane flux of solutes and consequently assigning TCs as having key roles in plant growth and productivity. However, TCs are typically positioned deep within tissues and have been studied mostly by electron microscopy. Recent advances in fluorophore labelling of plant cell walls using a modified pseudo-Schiff-propidium iodide (mPS-PI) staining procedure in combination with high-resolution Confocal microscopy have allowed visualization of cellular details of individual tissue layers in whole mounts, hence enabling study of tissue and cellular architecture without the need for tissue sectioning. Here we apply a simplified version of the mPS-PI procedure for Confocal Imaging of cellulose-enriched wall ingrowths in vascular TCs at the whole tissue level. The simplified mPS-PI staining procedure produced high-resolution three-dimensional images of individual cell types in vascular bundles and, importantly, wall ingrowths in phloem parenchyma (PP) TCs in minor veins of Arabidopsis leaves and companion cell TCs in pea. More efficient staining of tissues was obtained by replacing complex clearing procedures with a simple post-fixation bleaching step. We used this modified procedure to survey the presence of PP TCs in other tissues of Arabidopsis including cotyledons, cauline leaves and sepals. This high-resolution Imaging enabled us to classify different stages of wall ingrowth development in Arabidopsis leaves, hence enabling semi-quantitative assessment of the extent of wall ingrowth deposition in PP TCs at the whole leaf level. Finally, we conducted a defoliation experiment as an example of using this approach to statistically analyze responses of PP TC development to leaf ablation. Use of a modified mPS-PI staining technique resulted in high-resolution Confocal Imaging of polarized wall ingrowth deposition in TCs. This technique can be used in place of conventional electron microscopy and opens new possibilities to study mechanisms determining polarized deposition of wall ingrowths and use reverse genetics to identify regulatory genes controlling TC trans-differentiation.
Suong T T Nguyen - One of the best experts on this subject based on the ideXlab platform.
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high resolution Confocal Imaging of wall ingrowth deposition in plant transfer cells semi quantitative analysis of phloem parenchyma transfer cell development in leaf minor veins of arabidopsis
BMC Plant Biology, 2015Co-Authors: Suong T T Nguyen, David W MccurdyAbstract:Transfer cells (TCs) are trans-differentiated versions of existing cell types designed to facilitate enhanced membrane transport of nutrients at symplasmic/apoplasmic interfaces. This transport capacity is conferred by intricate wall ingrowths deposited secondarily on the inner face of the primary cell wall, hence promoting the potential trans-membrane flux of solutes and consequently assigning TCs as having key roles in plant growth and productivity. However, TCs are typically positioned deep within tissues and have been studied mostly by electron microscopy. Recent advances in fluorophore labelling of plant cell walls using a modified pseudo-Schiff-propidium iodide (mPS-PI) staining procedure in combination with high-resolution Confocal microscopy have allowed visualization of cellular details of individual tissue layers in whole mounts, hence enabling study of tissue and cellular architecture without the need for tissue sectioning. Here we apply a simplified version of the mPS-PI procedure for Confocal Imaging of cellulose-enriched wall ingrowths in vascular TCs at the whole tissue level. The simplified mPS-PI staining procedure produced high-resolution three-dimensional images of individual cell types in vascular bundles and, importantly, wall ingrowths in phloem parenchyma (PP) TCs in minor veins of Arabidopsis leaves and companion cell TCs in pea. More efficient staining of tissues was obtained by replacing complex clearing procedures with a simple post-fixation bleaching step. We used this modified procedure to survey the presence of PP TCs in other tissues of Arabidopsis including cotyledons, cauline leaves and sepals. This high-resolution Imaging enabled us to classify different stages of wall ingrowth development in Arabidopsis leaves, hence enabling semi-quantitative assessment of the extent of wall ingrowth deposition in PP TCs at the whole leaf level. Finally, we conducted a defoliation experiment as an example of using this approach to statistically analyze responses of PP TC development to leaf ablation. Use of a modified mPS-PI staining technique resulted in high-resolution Confocal Imaging of polarized wall ingrowth deposition in TCs. This technique can be used in place of conventional electron microscopy and opens new possibilities to study mechanisms determining polarized deposition of wall ingrowths and use reverse genetics to identify regulatory genes controlling TC trans-differentiation.
Ralf H Adams - One of the best experts on this subject based on the ideXlab platform.
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sample preparation for high resolution 3d Confocal Imaging of mouse skeletal tissue
Nature Protocols, 2015Co-Authors: Anjali P Kusumbe, Saravana K Ramasamy, Andrea Starsichova, Ralf H AdamsAbstract:High-resolution Confocal Imaging is a vital tool for analyzing the 3D architecture and detailed spatial distribution of cells in situ. However, Imaging of skeletal tissue has remained technically challenging because of its calcified nature. Here we describe a protocol that allows high-resolution Imaging of skeletal tissue with preservation of cellular morphology and tissue architecture. The procedure involves tissue fixation, decalcification and cryosectioning of the mouse skeletal tissue to generate thick sections. The thick sections generated by this procedure are not only compatible with the analysis of genetically expressed fluorescent proteins but they also preserve antigenicity, thus enabling diverse combinations of antibody labeling. Further, this procedure also permits other fluorescence techniques such as TUNEL and ethynyl deoxyuridine (EdU) incorporation assays. Images resulting from the Confocal Imaging can be assessed qualitatively and quantitatively to analyze various parameters such as distribution and interrelationships of cell types. The technique is straightforward and robust, highly reproducible and can be completed in ∼11 d.
Alfred J Meixner - One of the best experts on this subject based on the ideXlab platform.
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one pot synthesis of micron partly hollow anisotropic dumbbell shaped silica core shell particles
Chemical Communications, 2016Co-Authors: Johannes Maisch, Alfred J Meixner, Farhad Jafarli, Thomas Chasse, Felix Blendinger, Alexander Konrad, Michael Metzger, Marc Brecht, L DahneAbstract:A facile method is described to prepare micron partly hollow dumbbell silica particles in a single step. The obtained particles consist of a large dense part and a small hollow lobe. The spherical dense core as well as the hollow lobe are covered by mesoporous channels. In the case of the smaller lobe these channels are responsible for the permeability of the shell which was demonstrated by Confocal Imaging and spectroscopy.
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a high numerical aperture parabolic mirror as Imaging device for Confocal microscopy
Optics Express, 2001Co-Authors: M A Lieb, Alfred J MeixnerAbstract:We explore the diffraction limited focusing and Confocal Imaging properties of a high NA parabolic mirror for Confocal Imaging and spectroscopy of nanoparticles and single molecules. Vector field calculations of the electric fields near focus for both linear and radially polarized illumination are discussed and show that the optical field can be similar tightly focused as in the case of a high NA objective lens. Furthermore they show that a high NA parabolic mirror allows an easy orientation of the polarization of the illuminating light in all spatial directions. The simulation of Confocal Imaging of single molecules is discussed and yields, that the use of radially polarized excitation light gives an easy access to their orientations.
Anjali P Kusumbe - One of the best experts on this subject based on the ideXlab platform.
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sample preparation for high resolution 3d Confocal Imaging of mouse skeletal tissue
Nature Protocols, 2015Co-Authors: Anjali P Kusumbe, Saravana K Ramasamy, Andrea Starsichova, Ralf H AdamsAbstract:High-resolution Confocal Imaging is a vital tool for analyzing the 3D architecture and detailed spatial distribution of cells in situ. However, Imaging of skeletal tissue has remained technically challenging because of its calcified nature. Here we describe a protocol that allows high-resolution Imaging of skeletal tissue with preservation of cellular morphology and tissue architecture. The procedure involves tissue fixation, decalcification and cryosectioning of the mouse skeletal tissue to generate thick sections. The thick sections generated by this procedure are not only compatible with the analysis of genetically expressed fluorescent proteins but they also preserve antigenicity, thus enabling diverse combinations of antibody labeling. Further, this procedure also permits other fluorescence techniques such as TUNEL and ethynyl deoxyuridine (EdU) incorporation assays. Images resulting from the Confocal Imaging can be assessed qualitatively and quantitatively to analyze various parameters such as distribution and interrelationships of cell types. The technique is straightforward and robust, highly reproducible and can be completed in ∼11 d.