I am, however, clear-eyed and non-native enough to know that it's a disgrace that so much is concentrated here: no wonder the country has no idea what its own face looks like when the majority of its politics and media are fixed in one place.
After removal of culture media, cells were fixed with −10°C cold methanol.
After a controlled, 6-hr incubation in AHA-containing methionine-free media, neurons were fixed and AHA was fluorescently labeled.
After removing culture media, cells were fixed with 4% PFA, permeabilized in 0.5% Triton X100 in PBS and blocked with 2% BSA.
Following 2 days of culture in serum- and microsphere-supplemented media, cells were fixed using 10% buffered formalin, washed in PBS, and stained with 1 μg mL 1 DAPI (Invitrogen) in PBS.
G. vaginalis 14018 was grown to in culture media and bacterial cells were fixed on a glass chamber slide using 4% paraformaldehyde.
After incubation, media were removed; cells were fixed with methanol and stained with Diff Quick (Siemens Healthcare Diagnostics, IL, USA).
Injected embryos were cultured for 16 hr in KSOM media after which they were fixed for immunofluorescence analysis.
To visualize colonies contained 50 or more cells during the 14 days of culture, media was removed, cells were fixed in 3.7% paraformaldehyde for 15 min and stained with crystal violet and the colonies were counted under light microscope.
Briefly, upon removal of media, cells in each well were fixed with 200 μL of 10% cold TCA [Sigma-Aldrich, St Louis, Missouri, USA] (w/v; in deionised water).
