scholarly journals Cell biology of the future: Nanometer-scale cellular cartography

2015 ◽  
Vol 211 (2) ◽  
pp. 211-214 ◽  
Author(s):  
Justin W. Taraska
Keyword(s):  
Electron Microscopy ◽  
Cell Biology ◽  
Cellular Structure ◽  
Three Dimensional ◽  
Super Resolution ◽  
Molecular Organization ◽  
Nanometer Scale ◽  
Cellular Regulation ◽  
New Developments ◽  
Quantitative Image

Understanding cellular structure is key to understanding cellular regulation. New developments in super-resolution fluorescence imaging, electron microscopy, and quantitative image analysis methods are now providing some of the first three-dimensional dynamic maps of biomolecules at the nanometer scale. These new maps—comprehensive nanometer-scale cellular cartographies—will reveal how the molecular organization of cells influences their diverse and changeable activities.

scholarly journals Nanoscale subcellular architecture revealed by multicolor 3D salvaged fluorescence imaging

2019 ◽  
Author(s):  
Yongdeng Zhang ◽  
Lena K. Schroeder ◽  
Mark D. Lessard ◽  
Phylicia Kidd ◽  
Jeeyun Chung ◽  
...  
Keyword(s):  
Electron Microscopy ◽  
Cell Biology ◽  
Mammalian Cells ◽  
Spatial Organization ◽  
Protein Complexes ◽  
Three Dimensional ◽  
Super Resolution ◽  
Membrane Structures ◽  
Molecular Specificity ◽  
20 Nm

AbstractCombining the molecular specificity of fluorescent probes with three-dimensional (3D) imaging at nanoscale resolution is critical for investigating the spatial organization and interactions of cellular organelles and protein complexes. We present a super-resolution light microscope that enables simultaneous multicolor imaging of whole mammalian cells at ~20 nm 3D resolution. We show its power for cell biology research with fluorescence images that resolved the highly convoluted Golgi apparatus and the close contacts between the endoplasmic reticulum and the plasma membrane, structures that have traditionally been the imaging realm of electron microscopy.One Sentence SummaryComplex cellular structures previously only resolved by electron microscopy can now be imaged in multiple colors by 4Pi-SMS.

scholarly journals Three-Dimensional Measurements of Micro- to Nanometer-Scale Features at and Below the Surface Using Scanning Electron Microscopy

2011 ◽  
Vol 17 (S2) ◽  
pp. 992-993
Author(s):  
M Zhao ◽  
B Ming ◽  
P Kavuri ◽  
A Vladár
Keyword(s):  
Electron Microscopy ◽  
Scanning Electron Microscopy ◽  
Three Dimensional ◽  
Nanometer Scale ◽  
Dimensional Measurements ◽  
Scanning Electron

Extended abstract of a paper presented at Microscopy and Microanalysis 2011 in Nashville, Tennessee, USA, August 7–August 11, 2011.

scholarly journals Subaxolemmal cytoskeleton in squid giant axon. II. Morphological identification of microtubule- and microfilament-associated domains of axolemma.

1986 ◽  
Vol 102 (5) ◽  
pp. 1710-1725 ◽  
Author(s):  
S Tsukita ◽  
S Tsukita ◽  
T Kobayashi ◽  
G Matsumoto
Keyword(s):  
Electron Microscopy ◽  
Three Dimensional ◽  
Preceding Paper ◽  
Microscopic Analysis ◽  
Giant Axon ◽  
Squid Giant Axon ◽  
Molecular Organization ◽  
Fixation Method ◽  
Morphological Identification

In the preceding paper (Kobayashi, T., S. Tsukita, S. Tsukita, Y. Yamamoto, and G. Matsumoto, 1986, J. Cell Biol., 102:1710-1725), we demonstrated biochemically that the subaxolemmal cytoskeleton of the squid giant axon was highly specialized and mainly composed of tubulin, actin, axolinin, and a 255-kD protein. In this paper, we analyzed morphologically the molecular organization of the subaxolemmal cytoskeleton in situ. For thin section electron microscopy, the subaxolemmal cytoskeleton was chemically fixed by the intraaxonal perfusion of the fixative containing tannic acid. With this fixation method, the ultrastructural integrity was well preserved. For freeze-etch replica electron microscopy, the intraaxonally perfused axon was opened and rapidly frozen by touching its inner surface against a cooled copper block (4 degrees K), thus permitting the direct stereoscopic observation of the cytoplasmic surface of the axolemma. Using these techniques, it became clear that the major constituents of the subaxolemmal cytoskeleton were microfilaments and microtubules. The microfilaments were observed to be associated with the axolemma through a specialized meshwork of thin strands, forming spot-like clusters just beneath the axolemma. These filaments were decorated with heavy meromyosin showing a characteristic arrowhead appearance. The microtubules were seen to run parallel to the axolemma and embedded in the fine three-dimensional meshwork of thin strands. In vitro observations of the aggregates of axolinin and immunoelectron microscopic analysis showed that this fine meshwork around microtubules mainly consisted of axolinin. Some microtubules grazed along the axolemma and associated laterally with it through slender strands. Therefore, we were led to conclude that the axolemma of the squid giant axon was specialized into two domains (microtubule- and microfilament-associated domains) by its underlying cytoskeletons.

scholarly journals Electron tomography of cells

2011 ◽  
Vol 45 (1) ◽  
pp. 27-56 ◽  
Author(s):  
Lu Gan ◽  
Grant J. Jensen
Keyword(s):  
Electron Microscope ◽  
Cell Biology ◽  
3D Imaging ◽  
Imaging Technique ◽  
Electron Tomography ◽  
Three Dimensional ◽  
Molecular Organization ◽  
Biological Structure ◽  
Structural Cell ◽  
Projection Images

AbstractThe electron microscope has contributed deep insights into biological structure since its invention nearly 80 years ago. Advances in instrumentation and methodology in recent decades have now enabled electron tomography to become the highest resolution three-dimensional (3D) imaging technique available for unique objects such as cells. Cells can be imaged either plastic-embedded or frozen-hydrated. Then the series of projection images are aligned and back-projected to generate a 3D reconstruction or ‘tomogram’. Here, we review how electron tomography has begun to reveal the molecular organization of cells and how the existing and upcoming technologies promise even greater insights into structural cell biology.

Fine structure and molecular organization of the periostracum in a gastropod mollusc Buccinum undatum L. and its relation to similar structural protein systems in other invertebrates

1978 ◽  
Vol 283 (998) ◽  
pp. 417-459 ◽  
Keyword(s):  
Electron Microscopy ◽  
Cholesteric Liquid Crystal ◽  
Three Dimensional ◽  
Coiled Coil ◽  
Longitudinal Axis ◽  
Molecular Organization ◽  
Structural Protein ◽  
Buccinum Undatum ◽  
Basic Features ◽  
Smooth Transitions

The horny layer of periostracum which covers the shell of Buccinum undatum L. has been studied by a combination of the fine structural techniques including high resolution transmission electron microscopy and scanning electron microscopy as well as by chemical analysis and X-ray diffraction. It has been found that the main structural component is a tectin type protein with globular and probably coiled-coil α-helical regions accompanied by a small amount of polysaccharide. Much of the periostracum is built up of protein sheets superposed in a regular manner and stabilized by some type of covalent cross-linking involving aromatic molecules. The protein is one of the class of structural macromolecules called scleroproteins. Each sheet of protein is made up of molecular sub-units which have a characteristic dumb-bell shape and which are about 32 nm long and 6.5 nm wide at their globular ends. End-to-end long-axis aggregation of these units produces filaments which aggregate further by side-to-side association into ribbons and ultimately sheets. The side-to-side association is always in register and hence the sheets have a major transverse striation repeating at 32 nm intervals. The protein sheets can be ascribed a longitudinal axis in terms of the direction of their component filaments. On this basis it can be shown that successive superposed sheets are rotated in a horizontal plane through an angle of 20-25° relative to one another, in a constant direction either clockwise or anticlockwise. Such helicoidal organization is of the cholesteric liquid crystal type which is often found in a biological context, e.g. chitin fibril disposition in arthropod cuticle. This helicoidal layering of the protein sheets is manifested in oblique sections of periostracum as repeated parabolic lamellae. Irregularities in the form of the parabolic lamellae can be accounted for on the basis of the curvature and extensive folding of the periostracum. The outer and innermost layers of the periostracum tend not to show helicoidal organization but exhibit a different aggregation mode of the dumb-bell-shaped units into a three-dimensional hexagonally packed network matrix. This matrix is much interrupted by vacuoles and localized smooth transitions into the ribbon mode of aggregation. This ability to exist in both fibrous and network aggregation states is comparable to that known among the collagens and muscle proteins. The amino acid compositions and conformations of proteins which can form cholesteric helicoidal systems are reviewed and compared with the protein of Buccinum periostracum. This property is apparently confined to alpha helical rod-shaped proteins and globular tektins. The beta conformation does not favour cholesteric organization. The structures and compositions of other molluscan periostraca and periostracum- like structures from other invertebrate phyla are compared with the periostracum of Buccinum . While all periostraca and functionally related structures have certain basic features in common there is a considerable degree of variation at the molecular and organizational levels.

Characterization of Nanometer-Scale Porosity in Reservoir Carbonate Rock by Focused Ion Beam–Scanning Electron Microscopy

2012 ◽  
Vol 18 (1) ◽  
pp. 171-178 ◽  
Author(s):  
Bijoyendra Bera ◽  
Naga Siva Kumar Gunda ◽  
Sushanta K. Mitra ◽  
Douglas Vick
Keyword(s):  
Electron Microscopy ◽  
Scanning Electron Microscopy ◽  
Carbonate Rock ◽  
Focused Ion Beam ◽  
Ion Beam ◽  
Three Dimensional ◽  
Nanometer Scale ◽  
Beam Scanning ◽  
Sedimentary Carbonate ◽  
Scanning Electron

AbstractSedimentary carbonate rocks are one of the principal porous structures in natural reservoirs of hydrocarbons such as crude oil and natural gas. Efficient hydrocarbon recovery requires an understanding of the carbonate pore structure, but the nature of sedimentary carbonate rock formation and the toughness of the material make proper analysis difficult. In this study, a novel preparation method was used on a dolomitic carbonate sample, and selected regions were then serially sectioned and imaged by focused ion beam–scanning electron microscopy. The resulting series of images were used to construct detailed three-dimensional representations of the microscopic pore spaces and analyze them quantitatively. We show for the first time the presence of nanometer-scale pores (50–300 nm) inside the solid dolomite matrix. We also show the degree of connectivity of these pores with micron-scale pores (2–5 μm) that were observed to further link with bulk pores outside the matrix.

scholarly journals Expansion microscopy facilitates quantitative super-resolution studies of cytoskeletal structures in kinetoplastid parasites

Open Biology ◽  
2021 ◽  
Vol 11 (9) ◽  
pp. 210131 ◽  
Author(s):  
Peter Gorilak ◽  
Martina Pružincová ◽  
Hana Vachova ◽  
Marie Olšinová ◽  
Marketa Schmidt Cernohorska ◽  
...  
Keyword(s):  
Mitotic Spindle ◽  
Cell Biology ◽  
Leishmania Major ◽  
Three Dimensional ◽  
Super Resolution ◽  
Cell Types ◽  
Three Dimensional Reconstruction ◽  
Resolution Method ◽  
Entire Volume ◽  
Dimensional Reconstruction

Expansion microscopy (ExM) has become a powerful super-resolution method in cell biology. It is a simple, yet robust approach, which does not require any instrumentation or reagents beyond those present in a standard microscopy facility. In this study, we used kinetoplastid parasites Trypanosoma brucei and Leishmania major , which possess a complex, yet well-defined microtubule-based cytoskeleton, to demonstrate that this method recapitulates faithfully morphology of structures as previously revealed by a combination of sophisticated electron microscopy (EM) approaches. Importantly, we also show that due to the rapidness of image acquisition and three-dimensional reconstruction of cellular volumes ExM is capable of complementing EM approaches by providing more quantitative data. This is demonstrated on examples of less well-appreciated microtubule structures, such as the neck microtubule of T. brucei or the pocket, cytosolic and multivesicular tubule-associated microtubules of L. major . We further demonstrate that ExM enables identifying cell types rare in a population, such as cells in mitosis and cytokinesis. Three-dimensional reconstruction of an entire volume of these cells provided details on the morphology of the mitotic spindle and the cleavage furrow. Finally, we show that established antibody markers of major cytoskeletal structures function well in ExM, which together with the ability to visualize proteins tagged with small epitope tags will facilitate studies of the kinetoplastid cytoskeleton.

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