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Biomedical subjects

S Lamont

Publications and source records attributed to S Lamont.

10 recordsLinked to original sources

Electron tomography of large, multicomponent biological structures.

Electron tomography is an extremely useful method for deriving three-dimensional structure from electron microscope images. The application of this technique to the reconstruction of large, complex structures such as mitochondria is described in conjunction with several tools for segmentation, measurement, classification, and visualization. In addition, the use of massively parallel computers to perform the tomographic reconstruction efficiently using R-weighted backprojection or iterative techniques is described.

Algorithms

Serial section electron tomography: a method for three-dimensional reconstruction of large structures.

We present a method for combining single axis tomography and serial sectioning techniques to derive a three-dimensional reconstruction of large structures at electron microscopic resolution. This serial-tomography method allows the use of sufficiently thin sections to achieve adequate resolution with electron tomography, yet enables the generation of large reconstructions with considerably fewer sections than would be required using a serial thin section reconstruction technique. Serial thick sections (1-2 microns) are cut through the structure of interest, tomographic volume reconstructions are obtained for each section from a single axis tilt series, and the resulting series of volumes are then aligned and combined to form a single large volume. The serial-tomography method is illustrated with several samples, including red blood cells, the Golgi apparatus, and a spiny dendrite of a cortical pyramidal neuron. In some of these samples, the reconstruction is compared to correlated light microscopic views. The resulting large volume reconstructions appear to represent accurately the size and shape of objects such as red blood cells and spiny dendrites. The continuity of complex, tortuous structures such as the Golgi apparatus is also maintained across serial volumes. These examples demonstrate that it is possible to align and link a series of tomographic volumes accurately and that serial-tomography is a useful method for reconstructing relatively large structures without resorting to large numbers of serial thin sections.

Animals

Re-evaluation of the structural organization of neuritic plaques in Alzheimer's disease.

We re-examined the relationship among synaptic pathology, subcellular abnormalities within neurites in the plaques and beta-amyloid deposits of Alzheimer's disease (AD) using laser confocal imaging and computer-aided serial section reconstruction techniques. Analysis of serial optical sections of neuritic plaques double-immunolabeled for anti-beta-amyloid/anti-tau-2 revealed that 35% of this type of plaque contained a dense amyloid core with clusters of peripheral abnormal neurites. The other 65% were without a dense core and were mainly composed of abundant abnormal neuritic clusters with bundles of amyloid distributed throughout the neuritic plaque. While two-dimensional (2-D) analysis of the plaques has suggested that neurites are distributed in the plaque periphery with beta-amyloid localized in its center, serial section analysis showed the opposite arrangement can also be true. Three-dimensional (3-D) reconstructions of serial optical sections showed that the tau-positive tortuous axons clustered in the neuritic plaques were often continuous with synaptophysin-positive distended terminals. Analysis of electron micrographs from serial sections showed continuity among the different segments of the neurites. Further analysis of the computer generated 3-D reconstructed neuritic plaques (both from serial electron micrographs and serial optical sections), viewed as continuous rotating loops, confirmed that a great majority of the plaque volume was occupied by the clustered and continuous abnormal neurites, while the amyloid fibrils were compressed and displaced to the periphery of the plaque. The 3-D imaging of the neuritic plaques in AD suggests a more widespread and active neuritic damage than that predicted from 2-D observations.(ABSTRACT TRUNCATED AT 250 WORDS)

Aged

Programs for visualization in three-dimensional microscopy.

Three-dimensional data representing biological structures can be derived using several methods, including serial section reconstruction, optical sectioning, and tomography. The investigation, comprehension, and communication of structural relationships to others is greatly facilitated by computer-based visualization procedures. We describe SYNU, a suite of programs developed for interactive investigation of three-dimensional structure and for the production of high-quality three-dimensional images and animations. We illustrate the capabilities of SYNU in applications to biological data obtained by confocal light microscopy, serial section, and high-resolution electron microscopy from investigations at the cellular, subcellular, and molecular levels.

Actin Cytoskeleton

Restriction fragment length polymorphism analysis of major histocompatibility complex class II genes from inbred chicken lines.

High molecular weight DNA was extracted from sperm from chickens of 14 inbred lines. The DNA was digested with each of four restriction enzymes (Pvu II, Hind III, Bgl II, and Bam HI), electrophoresed for 18 or 45 h, blotted onto nitrocellulose, and hybridized to a chicken major histocompatibility complex (MHC, B complex) class II beta-chain probe (beta 2-exon specific). Restriction fragment length polymorphisms (RFLPs) were found with each of the restriction enzymes used. Birds with the same B haplotype always showed the same RFLP pattern; however, some birds of different B haplotypes also shared the same RFLP pattern. To test for the Mendelian inheritance of the RFLP patterns, the F2 progeny of an informative cross were analysed. The RFLP patterns corresponded with the serologically determined B haplotypes of the F2 birds, thereby showing the Mendelian inheritance of the polymorphic bands.

Animals