Massive Accumulation of Strontium and Barium in Diplonemid Protists

Pilátová, Jana; Tashyreva, Daria; Týč, Jiří; Vancová, Marie; Bokhari, Syed Nadeem Hussain; Skoupý, Radim; Klementová, Mariana; Küpper, Hendrik; Mojzeš, Peter; Lukeš, Julius

doi:10.1128/mbio.03279-22

Cited by 6 publications

(1 citation statement)

References 64 publications

(126 reference statements)

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“…Especially, if electrons are scattered in low vacuum and only a fraction is detected [ 17 , 29 ]. For example, in our previous study on single cell diplonemids where the extra resin could not be avoided [ 38 ], and high details were required, electron dose 34.7 eV/nm 2 and (long) pixel dwell times 4 μs had to be used. A final pixel size 6 × 6 nm 2 in x, y was achieved, but the voxel thickness in z had to be sacrificed and 100 nm thick sections had to be cut to ensure reliable cutting.…”

Section: Discussionmentioning

confidence: 99%

Minimal resin embedding of SBF-SEM samples reduces charging and facilitates finding a surface-linked region of interest

Konopová,

Týč

2023

Front Zool

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Background For decoding the mechanism of how cells and organs function information on their ultrastructure is essential. High-resolution 3D imaging has revolutionized morphology. Serial block face scanning electron microscopy (SBF-SEM) offers non-laborious, automated imaging in 3D of up to ~ 1 mm3 large biological objects at nanometer-scale resolution. For many samples there are obstacles. Quality imaging is often hampered by charging effects, which originate in the nonconductive resin used for embedding. Especially, if the imaged region of interest (ROI) includes the surface of the sample and neighbours the empty resin, which insulates the object. This extra resin also obscures the sample’s morphology, thus making navigation to the ROI difficult. Results Using the example of small arthropods and a fish roe we describe a workflow to prepare samples for SBF-SEM using the minimal resin (MR) embedding method. We show that for imaging of surface structures this simple approach conveniently tackles and solves both of the two major problems—charging and ROI localization—that complicate imaging of SBF-SEM samples embedded in an excess of overlying resin. As the surface ROI is not masked by the resin, samples can be precisely trimmed before they are placed into the imaging chamber. The initial approaching step is fast and easy. No extra trimming inside the microscope is necessary. Importantly, charging is absent or greatly reduced meaning that imaging can be accomplished under good vacuum conditions, typically at the optimal high vacuum. This leads to better resolution, better signal to noise ratio, and faster image acquisition. Conclusions In MR embedded samples charging is minimized and ROI easily targeted. MR embedding does not require any special equipment or skills. It saves effort, microscope time and eventually leads to high quality data. Studies on surface-linked ROIs, or any samples normally surrounded by the excess of resin, would benefit from adopting the technique.

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Section: Discussionmentioning

confidence: 99%

Minimal resin embedding of SBF-SEM samples reduces charging and facilitates finding a surface-linked region of interest

Konopová,

Týč

2023

Front Zool

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Co-removal and recycling of Ba2+ and Ca2+ in hypersaline wastewater based on the microbially induced carbonate precipitation technique: Overlooked Ba2+ in extracellular and intracellular vaterite

Yan,

Zhu,

Liu

et al. 2024

Journal of Hazardous Materials

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Ultrastructure and 3D reconstruction of a diplonemid protist (Diplonemea) and its novel membranous organelle

Tashyreva,

Týč,

Horák

et al. 2023

mBio

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Although diplonemid protists (Diplonemea, Euglenozoa) are among the most species-rich microeukaryotes in the ocean, many ultrastructural features of this lineage remain to be clarified. Using serial block-face scanning and transmission electron microscopy, we provide the whole-cell three-dimensional model of a previously undescribed diplonemid Lacrimia vacuolata sp. nov. For the first time, we were able to reconstruct a diplonemid cell in detail, including all its organelles and the flagellar and feeding apparatuses. By analyzing various stages of the cell cycle, we provide the first description of a diplonemid cell division, which is characterized by the disassembly of flagellar axonemes and the feeding apparatus and their further construction in the daughter cells. Moreover, we have identified a novel, ultrastructurally complex organelle, herein named the colv ( C enter for O rganization of L ayered V esicles). We suggest that the colv is involved in food processing and membrane trafficking, and describe its close association with other components of the cellular digestive system. IMPORTANCE The knowledge of cell biology of a eukaryotic group is essential for correct interpretation of ecological and molecular data. Although diplonemid protists are one of the most species-rich lineages of marine eukaryotes, only very fragmentary information is available about the cellular architecture of this taxonomically diverse group. Here, a large serial block-face scanning electron microscopy data set complemented with light and fluorescence microscopy allowed the first detailed three-dimensional reconstruction of a diplonemid species. We describe numerous previously unknown peculiarities of the cellular architecture and cell division characteristic for diplonemid flagellates, and illustrate the obtained results with multiple three-dimensional models, comprehensible for non-specialists in protist ultrastructure.

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Massive Accumulation of Strontium and Barium in Diplonemid Protists

Cited by 6 publications

References 64 publications

Minimal resin embedding of SBF-SEM samples reduces charging and facilitates finding a surface-linked region of interest

Minimal resin embedding of SBF-SEM samples reduces charging and facilitates finding a surface-linked region of interest

Co-removal and recycling of Ba2+ and Ca2+ in hypersaline wastewater based on the microbially induced carbonate precipitation technique: Overlooked Ba2+ in extracellular and intracellular vaterite

Ultrastructure and 3D reconstruction of a diplonemid protist (Diplonemea) and its novel membranous organelle

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