Evo 15 scanning electron

At necropsy, samples from the nasal cavity (cranial and caudal conchae, septum) were collected for scanning electron microscopy (SEM). Samples were fixed in 5% glutaraldehyde buffered with 0.1 M cacodylate buffer (Serva Electrophoresis) and were subsequently embedded by a modified osmium (O)-thiocarbohydrazide (T)-embedding (OTOTO) protocol, followed by critical-point-drying and coating with gold in a sputter-coater (SCD040, Oerlikon Balzers), as described previously.¹⁰
Embedded samples were mounted on 0,5” Aluminum Specimen Stubs (Agar Scientific) using 12 mm Leit-Tabs (Plano) and Conductive Carbon Cement after Göcke (Plano) and examined using a Zeiss EVO 15 scanning electron microscope (Carl Zeiss Microscopy) operating with 10 kV.

Neutrophils were harvested, seeded on glass cover slips, and stimulated with either RPMI, MβCD, or Simvastatin as stated above. After 3 h incubation under either hypoxia or normoxia, cells were fixed in 1.5% glutaraldehyde (Sigma-Aldrich Chemie GmbH, St. Louis, MO, USA) and 3% PFA, buffered with 0.1 M cacodylate buffer (Serva Electrophoresis, Heidelberg, Germany) for 24 h and subsequently washed with 0.1 M cacodylate buffer. For further processing, the samples were embedded with 1% osmium tetroxide (Science Services GmbH, Munich, Germany) and dehydrated in a series of graded ethanol, followed by critical-point-drying and coating with gold in a sputter-coater (SCD040, Oerlikon Balzers), as described previously [44 (link),45 (link)] Afterward, the samples were mounted on 0.5” Aluminum Specimen Stubs (Agar Scientific, Stansted, Essex, UK) using 12 mm Leit-Tabs (Plano) and examined using a Zeiss EVO 15 scanning electron microscope (Carl Zeiss Microscopy, Oberkochen, Germany) operating with an acceleration voltage of 10 kV.