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Eyecups at 150 days post-differentiation were fixed in one half strength Karnovsky fixative as described previously (PMID 17591911), followed by osmication, dehydration, and embedment in Epon resin.
Cultures were incubated for 20 hr at 37°C in 5% CO2, and were then either snap frozen in liquid nitrogen or fixed in one half strength Karnovsky fixative.
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Digital images were collected with Zeiss Axiocam HR cameras using Zeiss Axiovision 3. Embryos were fixed in one-half Karnovsky's fixative overnight, postfixed with 1.0% osmium tetroxide and stained en bloc with uranyl acetate in cacodylate buffer.
One-half of the trials was presented with the stimuli above the fixation cross and the other half were presented below fixation, in order to prevent the subjects from keeping their gaze fixed in one position.
Stationary robots are fixed in one place.
In one half.
RPE monolayers were fixed in half strength Karnovsky's fixative for 24 h at 4C.
Biopsies were split, one half fixed in formaldehyde and the other half snap-frozen in liquid nitrogen.
'Now watch this,' he says, lips fixed in a reassuring half grin, eyes twinkling.
The wound samples were divided in half; one half was fixed in McDowell's fixative for 48 h under refrigeration (McDowell and Trump, 1976) for historesin processing for light microscopy and transmission electron microscopy (TEM).
At the end of the experiments, 29 or 30 days following the start of the treatment with inhibitors, rats were killed and the right kidneys were sagittally sectioned to use one half fixed in formalin for immunohistochemistry for localization CGL and CBS proteins in the renal tissues, and the other half was snap-frozen for eventual western blot analysis.
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