Cm12 stem transmission electron microscope

Cell cultures, as well as pAVL and hAVL samples, were fixed with 2.5% glutaraldehyde diluted in 25 mM sodium acetate/acetic acid buffer, pH 4.8, containing 0.05% phthalocyanine cuprolinic blue (Electron Microscopy Sciences, Hatfield, PA, USA) and 0.05 M magnesium chloride, overnight at room temperature under constant stirring. Following this, bAVIC cultures and AVL samples were: (i) post-fixed with 2% osmium tetraoxide (Agar Scientific, Stansted, Essex, UK), (ii) dehydrated with graded ethanol solutions, and (iii) embedded into Epon 812 resin or Epon-Araldite resin, respectively. Ultrathin sections were collected onto formvar-coated 2 × 1-mm-slot copper grids and contrasted with uranyl acetate and lead citrate. Observations and photographic recording were undertaken using a CM12 STEM transmission electron microscope (Philips, Eindhoven, The Netherlands).

Cells were fixed with a 25 mM sodium acetate/acetic acid buffer containing 2.5% glutaraldehyde, 0.05% cuprolinic blue (Electron Microscopy Sciences, Hatfield, PA, USA), and 0.05 M magnesium chloride overnight at room temperature, keeping Petri dishes on a rocking platform. Cells were then post-fixed with 2% osmium tetraoxide (Agar Scientific, Stansted, Essex, UK), dehydrated with graded ethanol solutions, and embedded into Epon 812 resin. Ultrathin sections were collected onto formvar-coated 2 × 1 mm slot copper grids and contrasted with uranyl acetate and lead citrate. Observations and photographic recording were made using a Philips CM12 STEM transmission electron microscope.