SIRIS: a high resolution scanning infrared camera for examining paintings

The new SIRIS (Scanning InfraRed Imaging System) camera developed at the National Gallery in London allows highresolution images of paintings to be made in the near infrared region (900–1700 nm). Images of 5000 × 5000 pixels are made by moving a 320 × 256 pixel InGaAs array across the focal plane of the camera using two orthogonal translation stages. The great advantages of this camera over scanning infrared devices are its relative portability and that image acquisition is comparatively rapid – a full 5000 × 5000 pixel image can be made in around 20 minutes. The paper describes the development of the mechanical, optical and electronic components of the camera, including the design of a new lens. The software routines used to control image capture and to assemble the individual 320 × 256 pixel frames into a seamless mosaic image are also mentioned. The optics of the SIRIS camera have been designed so that the camera can operate at a range of resolutions; from around 2.5 pixels per millimetre on large paintings of up to 2000 × 2000 mm to 10 pixels per millimetre on smaller paintings or details of paintings measuring 500 × 500 mm. The camera is primarily designed to examine underdrawings in paintings; preliminary results from test targets and paintings are presented and the quality of the images compared with those from other cameras currently used in this field

Bcl-xL-mediated remodeling of rod and cone synaptic mitochondria after postnatal lead exposure: electron microscopy, tomography and oxygen consumption.

PurposePostnatal lead exposure produces rod-selective and Bax-mediated apoptosis, decreased scotopic electroretinograms (ERGs), and scotopic and mesopic vision deficits in humans and/or experimental animals. Rod, but not cone, inner segment mitochondria were considered the primary site of action. However, photoreceptor synaptic mitochondria were not examined. Thus, our experiments investigated the structural and functional effects of environmentally relevant postnatal lead exposure on rod spherule and cone pedicle mitochondria and whether Bcl-xL overexpression provided neuroprotection.MethodsC57BL/6N mice pups were exposed to lead only during lactation via dams drinking water containing lead acetate. The blood [Pb] at weaning was 20.6±4.7 µg/dl, which decreased to the control value by 2 months. To assess synaptic mitochondrial structural differences and vulnerability to lead exposure, wild-type and transgenic mice overexpressing Bcl-xL in photoreceptors were used. Electron microscopy, three-dimensional electron tomography, and retinal and photoreceptor synaptic terminal oxygen consumption (QO(2)) studies were conducted in adult control, Bcl-xL, lead, and Bcl-xL/lead mice.ResultsThe spherule and pedicle mitochondria in lead-treated mice were swollen, and the cristae structure was markedly changed. In the lead-treated mice, the mitochondrial cristae surface area and volume (abundance: measure correlated with ATP (ATP) synthesis) were decreased in the spherules and increased in the pedicles. Pedicles also had an increased number of crista segments per volume. In the lead-treated mice, the number of segments/crista and fraction of cristae with multiple segments (branching) similarly increased in spherule and pedicle mitochondria. Lead-induced remodeling of spherule mitochondria produced smaller cristae with more branching, whereas pedicle mitochondria had larger cristae with more branching and increased crista junction (CJ) diameter. Lead decreased dark- and light-adapted photoreceptor and dark-adapted photoreceptor synaptic terminal QO(2). Bcl-xL partially blocked many of the lead-induced alterations relative to controls. However, spherules still had partially decreased abundance, whereas pedicles still had increased branching, increased crista segments per volume, and increased crista junction diameter. Moreover, photoreceptor and synaptic QO(2) were only partially recovered.ConclusionsThese findings reveal cellular and compartmental specific differences in the structure and vulnerability of rod and cone inner segment and synaptic mitochondria to postnatal lead exposure. Spherule and pedicle mitochondria in lead-exposed mice displayed complex and distinguishing patterns of cristae and matrix damage and remodeling consistent with studies showing that synaptic mitochondria are more sensitive to Ca(2+) overload, oxidative stress, and ATP loss than non-synaptic mitochondria. The lead-induced decreases in QO(2) likely resulted from the decreased spherule cristae abundance and smaller cristae, perhaps due to Bax-mediated effects as they occurred in apoptotic rod inner segments. The increase in pedicle cristae abundance and CJ diameter could have resulted from increased Drp1-mediated fission, as small mitochondrial fragments were observed. The mechanisms of Bcl-xL-mediated remodeling might occur via interaction with formation of CJ protein 1 (Fcj1), whereas the partial protection of synaptic QO(2) might result from the enhanced efficiency of energy metabolism via Bcl-xL's direct interaction with the F1F0 ATP synthase and/or regulation of cellular redox status. These lead-induced alterations in photoreceptor synaptic terminal mitochondria likely underlie the persistent scotopic and mesopic deficits in lead-exposed children, workers, and experimental animals. Our findings stress the clinical and scientific importance of examining synaptic dysfunction following injury or disease during development, and developing therapeutic treatments that prevent synaptic degeneration in retinal and neurodegenerative disorders even when apoptosis is blocked

A new camera for high-resolution infrared imaging of works of art

A new camera – SIRIS (scanning infrared imaging system) – developed at the National Gallery in London allows high-resolution images to be made in the near infrared region (900–1700 nm). The camera is based on a commercially available 320 × 256 pixel indium gallium arsenide area array sensor. This relatively small sensor is moved across the focal plane of the camera using two orthogonal translation stages to give images of c. 5000 × 5000 pixels. The main advantages of the SIRIS camera over scanning infrared devices or sequential image capture and mosaic assembly are its comparative portability and rapid image acquisition – making a 5000 × 5000 pixel image takes less than 20 minutes. The SIRIS camera can operate at a range of resolutions; from around 2.5 pixels per millimetre over an area of up to 2 × 2 m to 10 pixels per millimetre when examining an area measuring 0.5 × 0.5 m. The development of the mechanical, optical and electronic components of the camera, including the design of a new lens, is described. The software used to control image capture and to assemble the individual frames into a seamless mosaic image is mentioned. The camera was designed primarily to examine underdrawings in paintings; preliminary results from test targets and paintings imaged in situ are presented and the quality of the images compared with those from other cameras currently used for this application

Doctor of Philosophy

dissertationAll cells have distinct cellular architectures that are critical for their function. A dramatic example of this relationship can be observed in cells that undergo subcellular branching. In this case, cells must first specify distinct branch sites and then outgrow cellular projections from those sites, resulting in a branched cellular morphology. Branching morphology is a common type of cell shape, examples of which include glial oligodendrocytes found in the human brain, dendritic cells of the mammalian immune system and by far the best studied example, neurons. Tubulogenesis or lumen morphology is another type of cell formation common throughout biology. Cellular tubes transport liquids and gases within animal tissues and are often found in elaborate organ systems that span the entire body including the human respiratory and circulatory systems. Despite the importance of these forms of cell architecture, little is known about the genes and molecular machinery that are required for developing branched tubular cells. Drosophila larval tracheal terminal cells are single, highly branched cells that have a subcellular lumen running through each branch. These cells are located at the ends of a network of interconnected tubes and are the final, and critical step in delivery of oxygen and other gases to animal tissues. Terminal cell development, which occurs primarily during larval stages, includes three distinct morphological processes: cell growth, subcellular branching and tubulogenesis. Cell outgrowth is a general process that is used by many other cell types to enable the overall growth. Subcellular branching is a iv specialized process that includes sending cellular projections out from the plasma membrane towards other cellular targets. Lastly, tubulogenesis is the process of forming a space or lumen within a cell through which gas can flow. Here, we use Drosophila larval terminal cells, a component of the respiratory system, to investigate the cellular mechanisms required for development of two distinct cellular morphologies, subcellular branching morphogenesis and subcellular lumen formation. Work described here focuses primarily on progress made in elucidating mechanisms of branch specification and branch outgrowth. We have found that the PAR-polarity protein complex is required for terminal cell branching and that through the Rho GTPase Cdc42, and other PAR proteins, the exocyst facilitates polarized membrane addition required for terminal cell branch outgrowth

Detecting Repetitions and Periodicities in Proteins by Tiling the Structural Space

The notion of energy landscapes provides conceptual tools for understanding the complexities of protein folding and function. Energy Landscape Theory indicates that it is much easier to find sequences that satisfy the "Principle of Minimal Frustration" when the folded structure is symmetric (Wolynes, P. G. Symmetry and the Energy Landscapes of Biomolecules. Proc. Natl. Acad. Sci. U.S.A. 1996, 93, 14249-14255). Similarly, repeats and structural mosaics may be fundamentally related to landscapes with multiple embedded funnels. Here we present analytical tools to detect and compare structural repetitions in protein molecules. By an exhaustive analysis of the distribution of structural repeats using a robust metric we define those portions of a protein molecule that best describe the overall structure as a tessellation of basic units. The patterns produced by such tessellations provide intuitive representations of the repeating regions and their association towards higher order arrangements. We find that some protein architectures can be described as nearly periodic, while in others clear separations between repetitions exist. Since the method is independent of amino acid sequence information we can identify structural units that can be encoded by a variety of distinct amino acid sequences

In Vivo and Ex Vivo Confocal Microscopy for Dermatologic and Mohs Surgeons

Confocal microscopy is a modern imaging device that has been extensively applied in skin oncology. More specifically, for tumor margin assessment, it has been used in two modalities: reflectance mode (in vivo on skin patient) and fluorescence mode (on freshly excised specimen). Although in vivo reflectance confocal microscopy is an add-on tool for lentigo maligna mapping, fluorescence confocal microscopy is far superior for basal cell carcinoma and squamous cell carcinoma margin assessment in the Mohs setting. This article provides a comprehensive overview of the use of confocal microscopy for skin cancer margin evaluation

Endomycorrhizae in Miocene paleosols: Implications in biotite weathering and accumulation of dolomite in plant roots (SW Madrid Basin, Spain)

This work provides evidence of arbuscular mycorrhizae (or endomycorrizhae) and their role in mineral weathering and soil carbonate production from Miocene paleosols. Formation of the paleosols-bearing mycorrhizae took place in marginal lacustrine and distal-fan deposits in the Madrid Basin. Endomycorrizhae fossils are preserved as carbonate in biotite-filled rhizoliths. Fossilized mycorrhizae are morphologically identical to those of living counterparts and consist of two linked parts. The extraradicular segment surrounding the root is represented by a calcitic mycelium comprising spores and two types of hyphae that are seen to attach and to corrode the mineral grains. The intraradicular part is made of a network of branched filaments, hyphal coils, arbuscules and likely vesicles that are mixed with the cortical root cells of the plants fossilized as dolomicrosparite aggregates. The preferential formation of the dolomite aggregates on biotite grains in rhizoliths is interpreted to reflect a focused interest of the plants on this mineral and indicates the mineral may have been used as a source of plant nutrients. The close association of the carbonate with the silicate further suggests that the precipitation of dolomite in the root cells and the weathering of the biotite resulted from interrelated processes. Main mechanisms to produce mineral dissolution include organic acids and chelate secretions by the plant roots and fungi which would remove nutrients. These results confirm that carbonate paleosols provide an excellent reservoir of information on ecological interactions and biogeochemical cycling, and expand the range of biogenic processes and their resultant products involved in the formation of carbonate soils. This approach has broad applications given the abundance of carbonate paleosols and the endomycorrhiza record dating back 400 million years

A Morphological and Ultrastructural Study of the Anterior Digestive Tract of Adult Nile Tilapia Oreochromis niloticus

Among the most-used fish species in aquaculture is the Nile tilapia, due to its rapid growth rate and its adaptation to a wide range of farming conditions. A careful description of the morphology of the digestive tract, particularly the esophagus and stomach, allows a better understanding of the relationship between structure and function. Combining scanning and light microscopy we highlighted the presence of five different zones in the stomach (1: esophagus-gastric lumen passage; 2: descending glandular portion; 3: fundic portion; 4: ascending glandular portion; 5: gastric-pyloric transition portion). Histochemical investigation showed a secretion of carboxylates mucopolysaccharides along the esophagus and sulphated complex carbohydrates in the stomach. These results suggest that mucins play a protective role of the epithelial lining, which is essential for a correct digestive process. Finally, the characterization of the main cellular structures may be inspiring for more advanced studies aiming to decipher the role of specific molecules, such as neuropeptides, involved in the physiological digestive process