Also, multiple aggregates can be seen in many of the cells' cytoplasm, indicating uptake of the hybridized NPs, but this would need to be verified with ultramicrotome
sectioning and will be addressed in future studies.
Samples were prepared employing a cryogenic ultramicrotome
Leica Ultracut at -125[degrees]C and a cut rate of 2mm/min.
After treatment with epoxy resin ("Serva" Switzerland), ultrathin sections were cut using an LKB-III ultramicrotome
(Sweden), which were stained with uranyl acetate for 10 min and lead citrate for 10 min.
Blocks were sectioned at 1 [micro]m using a Reichert Cut S ultramicrotome
(Leica, Wien, Austria).
Then, the embedded FP was sliced into 70 nm thickness in longitudinal and transverse section with Leica UC7 ultramicrotome
. Finally, the slices were stained with uranyl acetate-lead citrate solution and observed by Hitachi H-7650 TEM with acceleration voltage of 100 kV.
Samples were sectioned using an ultramicrotome
, stained with uranyl acetate and lead citrate, and examined under an FEI Tecnai G2 electron microscope (FEI, Eindhoven, The Netherlands).
Ultra-thin sections were obtained using diamond knives with a Leica EM UC6 ultramicrotome
(Leica Co., Vienna, Austria).
The SFTSV-infected Vero cell monolayer was fixed according to described methods (11) and cut on ultramicrotome
(RMC MT-XL) at 65 nm.
Part of the material was embedded in Epon-Araldite mixture to prepare blocks with further making of ultramicrotome
The embedded samples were sectioned at 500 nm thick with a Power Tome PC (RMC Products) Ultramicrotome
, dyed with Touluidin Blue and observed with an Olympus IX-51 inverted light microscope.
After finding desired area ultrathin sections (~70 nm-thick) were cut by ultramicrotome
(ultracut UCT) with glass knives (Leica EMKMR2) at a thickness of silver-gold interface color and picked up on 100 mesh copper grids.
Semi-thin sections were obtained using an ultramicrotome
(Leica EM UC6, Austria).