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Cryo-SEM of cell cortex architecture and membrane ultrastructure in eugregarines

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VAŠKOVICOVÁ Naděžda KOVÁČIKOVÁ Magdaléna BARDŮNEK VALIGUROVÁ Andrea DIAKIN Andrei KRZYŽÁNEK Vladislav

Rok publikování 2018
Druh Konferenční abstrakty
Fakulta / Pracoviště MU

Přírodovědecká fakulta

Citace
Popis Cryo-SEM of frozen microscopic structures, especially unicellular organisms, is a fast method for processing the sample for electron microscopy. Cells suspensions in buffer were frozen in device HPF 100 (Leica microsystems) on 3 mm carriers and placed inside ACE 600 (Leica microsystems). Cells were fractured and after short etching transferred into Magellan 400 (FEI) by vacuum shuttle VCT100 (Leica microsystems) on cryo-holder. The cryo-SEM analysis was performed at temperature of -120°C, electron probe current 3.1 - 6.3 pA, and electron energy 1-2 keV. Freeze-fracture of cells and etching revealed organisation of filamentous structures and distribution of cytoplasmic organelles, mostly lipid drops, secretory vesicles and amylopectin granules. The cell cortex of gregarines with typical folded arrangement of the plasma membrane and both cytomembranes can be seen in cross fracture along with micropores, medium-sized pores localised in the lateral part of epicytic folds and ductus incorporated into cytomembranes.

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