Sugars were extracted with four successive 250 l aliquots of 80% (v/v) ice-cold ethanol and further grinding.
One-half was placed in oxygenated ice-cold superfusate and gently agitated in the dark.
Tissue was homogenized immediately in ice-cold 85% methanol to ensure protein inactivation.
The supernatant was discarded and the pellet washed once with 0.5 ml of ice-cold 75% ethanol.
For better preservation of cellular structure, the eye was first fixed in ice-cold 4% paraformaldehyde for 20 - 60 min before dissection.
The tubes were kept on ice for 5 min before the addition of 100 l of ice-cold 1:25 assay mix.
The ice-cold water, usually unpleasant, felt good!
Take away the rhetorical flourishes and polite formularies, and your speech today is an ice-cold, nationalist, declaration of war on the idea that that flag symbolises.
