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2009 Keynote speakers.
Wednesday 26th August -- Friday 28th August, 2009.

The programme abstract book can now be downloaded from here.

State of the art light and electron microscopy for cell biology.
Prof. Gareth Griffiths, Dept Molecular Biosciences, University of Oslo, Norway.

In the first part of this presentation I will cover the main EM techniques that are important for molecular cell biology.  An important conceptual breakthrough in EM was the introduction of cryo EM and the concept of vitrification by Jacques Dubochet and colleagues in the early 1980’s. This approach made it possible to avoid fixatives and other chemicals that had long been necessary for most EM methods. Two different cryo EM approaches have been developed, namely single particle methods and a sectioning method referred to as Cryo EM of Vitrified Sections (CEMOVIS). I will show examples of these methods, as well as briefly cover the method of tomography, that is being increasingly used in conjunction with cryo EM, and with conventional plastic embedding to provide three-dimensional models of biological specimens. I will also cover the importance of using light microscopy (LM) in conjunction with EM for cell biology. The use of both methods together is especially important for interpreting the results of immuno-labeling studies, an approach that still require the use of chemical fixatives in almost all cases. In the second part of the presentation I will focus on the use of LM and EM in our recent work on phagocytosis, addressing two systems, model latex beads and Mycobacteria, including M.tuberculosis. For LM live cell video microscopy will be described in order to reveal striking dynamic process of transient assembly of actin by latex bead phagosomes in GFP-actin expressing mouse macrophages. The subsequent part will address the use of different EM sectioning approaches to visualize different mycobacteria in macrophage phagosomes and will conclude that the best sectioning method to monitor the native structure of free mycobacteria and mycobacteria within phaagosomes is CEMOVIS.

Genome-wide imaging and analysis of membrane traffic pathways.
Jeremy C. Simpson, University College Dublin, Ireland

Mammalian cells have a highly complex internal architecture consisting of membrane-bounded organelles each with defined functions. Organelles within the secretory and endocytic pathways however do not function in isolation, but are able to exchange material through the highly dynamic process of membrane traffic. A more complete inventory and understanding of the membrane traffic machinery is an essential prerequisite to improved drug design and targeting. We have applied high-throughput systematic subcellular localisation, genome-wide gene downregulation by RNA interference, and automated high content screening microscopy as approaches to more comprehensively discover the protein machinery involved in cellular trafficking pathways. These experiments have revealed a wide diversity of previously unidentified membrane traffic regulators, and an unexpected high degree of inter-dependence with basic cellular processes.

Foundation principles of the interaction of nanoparticles with cells.
Anna Salvati, Iseult Lynch, and Kenneth Dawson, Centre for BioNano Interactions, University College Dublin, Belfield, Dublin 4, Ireland.

The importance of understanding the interactions between nanoscale materials and living matter has begun to be appreciated in recent years. Thanks to their size (10-100nm), nanoparticles give the opportunity to identify and study specific interactions both at cellular and at tissue level.
The underlying rationale is both real and durable. Nanoparticles of less than 100nm can enter cells and those of less than 35 nm can pass the blood brain barrier. These are fundamental size scales of biological relevance that will ensure that engineered nanoscience will impinge on biology and medicine for many decades. Likely, the uptake and trafficking of nanoparticles in contact with biological fluids is greatly affected by the proteins and other biomolecules that associate to their surface, and the protein corona taken from the surrounding biological milieu will constitute the real identity of nanoparticles exposed to cells. The pace of advance is extraordinary. This arena of research not only opens up new directions in nanomedicine and nanodiagnostics, but offers the chance to implement nanotechnology in a safe and responsible manner, addressing the concerns of nanosafety in parallel with the development of applications.
Fundamental in this scenario is the ability to follow (and ultimately control) the localisation in time and space of nanoparticles inside cells, in a quantitative and reproducible manner. This has been achieved by the combination of electron microscopy and fluorescence based techniques, including fluorescence microscopy and live cell imaging of single nanoparticles inside the cell, thus enabling us to study nanoparticle uptake from the early entry to their final localisation.

 

Advanced Imaging Approaches in Nanomedicine.
Yuri Volkov, Department of Clinical Medicine, Trinity College Dublin, Ireland.

Rapid development of nanotechnology consistently increases the likelihood of human contact with environmentally presented and engineered nanomaterials, i.e. the tiny objects ranging in size from one to several hundreds of nanometres and featuring an extreme diversity in shapes and physico-chemical properties (inorganic silica, carbon,  metal oxide, inert metal nanoparticles and core/shell structures, polymeric particles and spheres, nanorods, nanotubes, nanowires, dendrimers, liposomes and complex derivatives of all the above, to name but a few). Phagocytes, epithelium in the lungs and gastrointestinal tract as well as cells of the cardiovascular system are the primary candidates to encounter these nanomaterials in real life situations. Such encounters may cause specific functional responses, including triggering of the intracellular signaling cascades and immune reactions, apoptotic and direct toxic effects.  However, there is still very little definitive systematic information about the consequences of interactions of nano-scale objects with human cells and tissues of diverse origin and therefore safety-related issues are high on the agenda in the emerging scientific area of nanomedicine. On the other hand, optimistic expectations are associated with the opportunities of using the nanoparticles as a new class of drug delivery systems, arising from the fact that the finite, but tunable size of the engineered nanostructures used as drug delivery vehicles can impose very precise nano-scale drug distribution barriers at the level of cells, tissues and entire organism thereby eliminating undesirable side effects pertinent to most contemporary medicines. High content imaging and analysis approach in combination with live cell confocal imaging, atomic force microscopy, fluorescence lifetime imaging and Raman spectrometry provides a unique integrated technological toolkit for visualization and physical characterization of nanoparticle-cell interactions at the levels of uptake, intracellular transport, subcellular and organelle targeting of nanoparticles. An exceptional aspect of this approach is that it is possible to identify individual cell, as well as population responses associated with nanoparticle exposure, as in this way subtle effects on small groups of cells within the whole, which could be averaged out by routine screening, are fully registered and elucidated. We will provide here an overview of such multi-platform cell and subcellular imaging application scenarios for investigations of safety and intracellular distribution of nanomaterials with promising biomedical application potential in live human phagocytes and cells of non-phagocytic origin.
Supported by the Health Research Board of Ireland, Science Foundation of Ireland SRC BioNanoInteract and EU FP-6 Consortium NanoInteract.

High resolution electron microscopy applied to nanostructured oxides – structure function relationships and characterization.

C. O’Dwyer, Department of Physics, and Materials & Surface Science Institute
University of Limerick, Limerick, Ireland.

One-dimensional nanomaterials, such as nanotubes, nanowires, and nanobelts or nanoribbons have attracted considerable attention in the past decade because of their novel and useful physical properties leading to numerous applications. Although the majority of research and development has been based on carbonaceous and compound semiconducting nanostructures, attention is now being directed to transition metal nanostructures based on their oxides which, due to their versatile chemical properties often modulable by changes in the oxidation state in the metal co-ordination sphere, can lead to a variety of products and tuneable materials. Incorporating such nanostructures into known device configurations could improve on current designs while potentially allowing further functionality by exploiting nanoscale electronic and photonic properties of suitable materials.
This talk will outline recent findings by our group concerning the structure-function relationships of a range of metal oxide nanostructures: vanadium oxide, zinc oxide, and indium tin oxide. Starting from the laminar V2O5 xerogel numerous two-dimensional organic-inorganic VOx intercalation products have been obtained. Many of these nanostructures may be obtained in quantities on the order of grams. Structure related properties such as scrolling to form nanotubes, nanoscale actuation, charge storage for batteries and self-assembly will be addressed.
In parallel, nanostructures designed for optical emission and absorptive charge storage have garnered obvious interest in recent times, resulting from a drive to research new charge storage architectures and advanced solar cell designs. In this section, both ZnO and ITO nanowires will be shown to exhibit excellent optical properties. TEM was used in all cases to determine the nanostructure phase and relation to opto-electronic characteristics. ITO nanowires, grown as branched layers, will be shown to be fully transparent and conductive contact layers, optimized for application as fully transparent contacts in the visible to near-infra red region for silicon-based light emitting devices (LEDs).
For ZnO, this talk will outline how high resolution microscopy was used to probe the structure and growth of the Al-coated ZnO nanowires and related that information to their optical properties. These structures were grown by catalyst-free chemical vapour deposition (CVD). After removal from the growth substrate by sonication, a second mirror facet was obtained as an effect of the cleavage along the (0001) crystallographic plane, thus leading to an enhanced axial Fabry-Perot resonator. The resulting nanostructures form the lowest threshold core-shell UV nanolasers reported to date.

 

 

Updated 27-June-2009.