Temcam f416 r camera

Specimen grids were examined with a JEM 1400 transmission electron microscope (JEOL Co., Tokyo, Japan, 2008), equipped with a MORADA G2 11‐megapixel TEM camera (EMSIS GmbH, Münster, Germany) at Nencki Institute of Experimental Biology PAS or with a Tecnai T12 BioTwin electron microscope (FEI, Hillsboro, OR, USA) equipped with a 16 megapixel TemCam‐F416 (R) camera (TVIPS GmbH) at the International Institute of Molecular and Cell Biology in Warsaw.

RKO cells were seeded on Nunc Thermanox coverslips (150067; Thermo Fisher Scientific) on a 24-well plate. Furthermore, 24 h after seeding, cells were transfected with 30 nM siRNAs. Then 72 h after transfection, cells were fixed in 2.5% glutaraldehyde for 2 h, then washed three times with PBS, postfixed with 1% osmium tetroxide for 1 h, washed with water, and incubated in 1% aqueous uranyl acetate overnight at 4°C. Then the cells were dehydrated with increasing dilutions of ethanol, infiltrated with epoxy resin (45-359-1EA-F; Sigma-Aldrich), embedded using BEEM capsules (Hanson et al, 2010 (link)), and incubated at 60°C for 72 h. Then polymerized blocks were trimmed and cut with a Leica ultramicrotome (EM UC7) for ultrathin sections (65 nm thick) and collected on copper grids, mesh 300 (AGG2300C; Agar Scientific). Specimen grids were examined with a transmission electron microscope Tecnai T12 BioTwin (FEI) equipped with a 16-megapixel TemCam-F416 (R) camera (TVIPS GmbH).

Fresh samples of the spleen, about 3 square mm, were fixed in 2.5% glutaraldehyde for 24 hr at 4 ° C, then washed in PBS and postfixed with 1% osmium tetroxide for 1 hr. After washing with water they were incubated with 1% aqueous uranyl acetate for 12 hr at 4 ° C. Next, samples were dehydrated at room temperature with increasing concentrations of ethanol, infiltrated with epoxy resin (Sigma-Aldrich, 45-359-1EA-F) and subjected for polymerization for 48 hr at 60 ° C. Polymerized resin blocks were trimmed with a tissue processor (Leica EM TP), cut with an ultramicrotome (EM UC7, Leica) for ultrathin sections (65 nm thick), and collected on nickel grids, mesh 200 (Agar Scientific, G2200N). Specimen grids were examined with a transmission electron microscope Tecnai T12 BioTwin (FEI, Hillsboro, OR, USA) equipped with a 16 megapixel TemCam-F416 (R) camera (TVIPS GmbH) at in-house Microscopy and Cytometry Facility.