Quorum ems 150t es

S. aureus Newman Δspa was grown in phenol-free RPMI medium (Gibco) at 37 °C overnight. The bacteria were washed two times with PBS and resuspended in 0.05% bovine serum albumin (BSA) in phenol-free RPMI to an OD₆₀₀ ~ 1. 500 μl of bacteria was mixed with 500 μl of NHS (BioIVT) or 500 μl 0.05% BSA in phenol-free RPMI. The samples were incubated at 37 °C for 8 h with shaking. The bacteria were then washed three times with PBS. For EM analyses, samples were fixed 1:1 with a 2x fixative [5% glutaraldehyde, 4% paraformaldehyde (PFA) in 0.2 M sodium cacodylate buffer] and dehydrated through a graded series of ethanol. Samples were critical point dried using liquid carbon dioxide in a Tousimis Samdri 795 Critical Point Drier, coated with carbon in a Quorum EMS 150T ES (Quorum Technologies Ltd) and examined in a Zeiss Supra Field Emission Scanning Electron Microscope (Carl Zeiss Microscopy), using an accelerating voltage of 8 KV.

HCT116 and Hke3 [control, irinotecan (2μM), reovirus (5MOI) and combination] cells were fixed in 2.5% glutaraldehyde, 0.1 M sodium Cacodylate, 0.2 M Sucrose, 5mM MgCl2 pH 7.4 and dehydrated through a graded series of ethanol, then critically point dried using liquid carbon dioxide in a Tousimis Samdri 795 Critical Point Drier (Rockville MD). They were next sputter coated with chromium in a Quorum EMS 150T ES (Quorum Technologies Ltd, United Kingdom) and examined in a Zeiss Supra Field Emission Scanning Electron Microscope (Carl Zeiss Microscopy, LLC North America), using an accelerating voltage of 3KV and observed at 1K, 5K and 10K magnification.

Tracheas from E18.5 WT or Tnrc6a^gt/gt embryonic lungs were dissected, fixed in 2.5% glutaraldehyde, 0.1 M sodium Cacodylate, 0.2 M Sucrose, 5 mM MgCl2 pH 7.4 and dehydrated through a graded series of ethanol dilutions. Samples were processed using liquid carbon dioxide in a Tousimis Samdri 795 Critical Point Drier (Rockville MD), sputter coated with chromium in a Quorum EMS 150 T ES (Quorum Technologies Ltd, United Kingdom) and examined in a Zeiss Supra Field Emission Scanning Electron Microscope (Carl Zeiss Microscopy, LLC North America), using an accelerating voltage of 2 kV. For quantitative analysis, lungs (n = 2) were examined and about 120 cells were counted for each sample.