scholarly journals Expansion Microscopy provides new insights into the cytoskeleton of malaria parasites including the conservation of a conoid

2020 ◽  
Author(s):  
Eloïse Bertiaux ◽  
Aurélia C Balestra ◽  
Lorène Bournonville ◽  
Mathieu Brochet ◽  
Paul Guichard ◽  
...  
Keyword(s):  
Electron Microscopy ◽  
Crucial Role ◽  
Reduced Form ◽  
Malaria Parasites ◽  
Whole Cells ◽  
Cytoskeleton Organization ◽  
Dynamic Assembly ◽  
Microtubule Structure

AbstractMalaria is caused by unicellular Plasmodium parasites. Plasmodium relies on diverse microtubule cytoskeletal structures for its reproduction, multiplication or dissemination. Due to the small size of this parasite, its cytoskeleton has been primarily observable by electron microscopy. Here, we demonstrate that the nanoscale cytoskeleton organization is within reach using ultrastructure expansion microscopy (U-ExM). In developing microgametocytes, U-ExM allows to monitor the dynamic assembly of axonemes and concomitant tubulin polyglutamylation in whole cells. In the invasive merozoite and ookinete forms, U-ExM unveils the subpellicular microtubule arrays that confer cell rigidity. In ookinete, we additionally identify an apical tubulin ring above the subpellicular microtubules that colocalises with markers of the conoid in related Apicomplexa parasites. This microtubule structure was presumed to be lost in Plasmodium despite its crucial role in both motility and invasion in most apicomplexans. Here, U-ExM reveals that a divergent and reduced form of the conoid is actually conserved in the Plasmodium genus.

scholarly journals Expansion microscopy provides new insights into the cytoskeleton of malaria parasites including the conservation of a conoid

PLoS Biology ◽  
2021 ◽  
Vol 19 (3) ◽  
pp. e3001020
Author(s):  
Eloïse Bertiaux ◽  
Aurélia C. Balestra ◽  
Lorène Bournonville ◽  
Vincent Louvel ◽  
Bohumil Maco ◽  
...  
Keyword(s):  
Electron Microscopy ◽  
Crucial Role ◽  
Reduced Form ◽  
Malaria Parasites ◽  
Whole Cells ◽  
Apicomplexan Parasites ◽  
Dynamic Assembly

Malaria is caused by unicellular Plasmodium parasites. Plasmodium relies on diverse microtubule cytoskeletal structures for its reproduction, multiplication, and dissemination. Due to the small size of this parasite, its cytoskeleton has been primarily observable by electron microscopy (EM). Here, we demonstrate that the nanoscale cytoskeleton organisation is within reach using ultrastructure expansion microscopy (U-ExM). In developing microgametocytes, U-ExM allows monitoring the dynamic assembly of axonemes and concomitant tubulin polyglutamylation in whole cells. In the invasive merozoite and ookinete forms, U-ExM unveils the diversity across Plasmodium stages and species of the subpellicular microtubule arrays that confer cell rigidity. In ookinetes, we additionally identify an apical tubulin ring (ATR) that colocalises with markers of the conoid in related apicomplexan parasites. This tubulin-containing structure was presumed to be lost in Plasmodium despite its crucial role in motility and invasion in other apicomplexans. Here, U-ExM reveals that a divergent and considerably reduced form of the conoid is actually conserved in Plasmodium species.

Progressive Mucinous Histiocytosis: Importance of Electron Microscopy to Confirm Diagnosis

2010 ◽  
Vol 14 (5) ◽  
pp. 245-248 ◽  
Author(s):  
Iman Hemmati ◽  
W. Alastair McLeod ◽  
Richard I. Crawford
Keyword(s):  
Electron Microscopy ◽  
Light Microscopy ◽  
Clinical Examination ◽  
Crucial Role ◽  
Langerhans Cell Histiocytosis ◽  
Langerhans Cell ◽  
History Of ◽  
Characteristic Features ◽  
Immunohistochemical Studies

Background: Progressive mucinous histiocytosis (PMH) is a benign, non-Langerhans cell histiocytosis with characteristic ultrastructural features that can be used for diagnosis. Once an important tool in dermatologic diagnosis, electron microscopy has been largely replaced by immunohistochemistry and immunofluorescence techniques today. However, electron microscopy occasionally still plays a crucial role in the diagnosis of dermatologic conditions. We report a case of PMH as an example of a dermatologic disorder that requires electron microscopy for its diagnosis. Methods: A 60-year-old woman presented to our clinic with a history of small, sharply demarcated, skin-colored papules ranging from 2 to 5 mm in diameter distributed over the arms, forearms, and dorsal hands. The results of light microscopy, immunohistochemical studies, and clinical examination were inconclusive. Another biopsy for electron microscopy showed the characteristic features of PMH. Conclusion: This case demonstrates that a dermatopathology service still needs to have access to electron microscopy for diagnostic purposes to successfully diagnose a small number of rare conditions.

scholarly journals Electron microscopy snapshots of single particles from single cells

2018 ◽  
Vol 294 (5) ◽  
pp. 1602-1608 ◽  
Author(s):  
Xiunan Yi ◽  
Eric J. Verbeke ◽  
Yiran Chang ◽  
Daniel J. Dickinson ◽  
David W. Taylor
Keyword(s):  
Electron Microscopy ◽  
Single Particle ◽  
Protein Complexes ◽  
Signal To Noise Ratio ◽  
Single Cells ◽  
Particle Analysis ◽  
Biological Macromolecules ◽  
Single Particle Analysis ◽  
Whole Cells ◽  
Potential Applications

Cryo-electron microscopy (cryo-EM) has become an indispensable tool for structural studies of biological macromolecules. Two additional predominant methods are available for studying the architectures of multiprotein complexes: 1) single-particle analysis of purified samples and 2) tomography of whole cells or cell sections. The former can produce high-resolution structures but is limited to highly purified samples, whereas the latter can capture proteins in their native state but has a low signal-to-noise ratio and yields lower-resolution structures. Here, we present a simple, adaptable method combining microfluidic single-cell extraction with single-particle analysis by EM to characterize protein complexes from individual Caenorhabditis elegans embryos. Using this approach, we uncover 3D structures of ribosomes directly from single embryo extracts. Moreover, we investigated structural dynamics during development by counting the number of ribosomes per polysome in early and late embryos. This approach has significant potential applications for counting protein complexes and studying protein architectures from single cells in developmental, evolutionary, and disease contexts.

An open-access volume electron microscopy atlas of whole cells and tissues

Nature ◽  
2021 ◽  
Author(s):  
C. Shan Xu ◽  
Song Pang ◽  
Gleb Shtengel ◽  
Andreas Müller ◽  
Alex T. Ritter ◽  
...  
Keyword(s):  
Electron Microscopy ◽  
Open Access ◽  
Whole Cells ◽  
Volume Electron Microscopy

Phase Contrast Enhancement with Phase Plates in Biological Electron Microscopy

2010 ◽  
Vol 18 (4) ◽  
pp. 10-13 ◽  
Author(s):  
Kuniaki Nagayama ◽  
Radostin Danev ◽  
Hideki Shigematsu ◽  
Naoki Hosogi ◽  
Yoshiyuki Fukuda ◽  
...  
Keyword(s):  
Thin Film ◽  
Electron Microscopy ◽  
Phase Contrast ◽  
Phase Plate ◽  
Electron Loss ◽  
Whole Cells ◽  
Transmission Electron ◽  
Different Types ◽  
Transparent Objects ◽  
Phase Plates

Theoretically, transmission electron microscopy (TEM) is compatible with three different types of phase plate: thin-film, electrostatic, and magnetic. However, designing functional phase plates has been an arduous process that has suffered from unavoidable technical obstacles such as phase-plate charging and difficulties associated with micro-fabrication of electrostatic and magnetic phase plates. This review discusses phase-contrast schemes that allow visualization of transparent objects with high contrast. Next it deals with recent studies on biological applications ranging from proteins and viruses to whole cells. Finally, future prospects for overcoming the problem of phase-plate charging and for designing the next generation of phase-plates to solve the problem of electron loss inherent in thin-film phase plates are discussed.

scholarly journals Publisher Correction: An open-access volume electron microscopy atlas of whole cells and tissues

Nature ◽  
2021 ◽  
Author(s):  
C. Shan Xu ◽  
Song Pang ◽  
Gleb Shtengel ◽  
Andreas Müller ◽  
Alex T. Ritter ◽  
...  
Keyword(s):  
Electron Microscopy ◽  
Open Access ◽  
Whole Cells ◽  
Volume Electron Microscopy

Specimen Observation at Large Tilt Angles with a Top-Entry Goniometer Stage

1974 ◽  
Vol 32 ◽  
pp. 558-559
Author(s):  
Mircea Fotino
Keyword(s):  
Electron Microscopy ◽  
Critical Point ◽  
Whole Cells ◽  
Morphological Studies ◽  
Controlled Conditions ◽  
Critical Point Drying ◽  
Cells Cultured

In one of the most resourceful applications of million-volt electron microscopy to biological problems, whole cells cultured directly on grids under controlled conditions and maintained unperturbed in vacuo through critical-point drying are viewed stereoscopically for morphological studies.Great advantage would thus be drawn in this regard from high tilting flexibility that would allow specimen observation under larger tilting angles yielding side stereo views, as it were, in addition to the more routine top views.

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