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As previously described (Yang et al., 2011), animals were deeply anesthetized with isofluorane, decapitated and the brains removed in ice cold sectioning solution.
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Cold section and hot section materials from military turbojet engines were chosen for testing.
The hearts were removed, immersed in cold saline solution, rinsed, then sectioned into 7-mm short-axis slices from base to apex.
The excised segment was flushed of any intestinal content with cold saline solution and opened along the mesenteric border in ice cold-oxygenated Ringer solution.
The second scenario, hypoxic damage, was countered in as far as that is possible by keeping the tissue in ice-cold oxygenated buffer solution during sectioning.
Donor hearts were flushed with cold preservation solution (University of Wisconsin [UW] or Custodiol) and stored in the same solution.
To prepare retinal slices for electrophysiological experiments, a section of the eyecup was placed vitreal side down on a piece of filter paper (2×5 mm, AAWP, 0.8 mm pores, Millipore, Bedford, MA, USA) and isolated in cold saline solution.
A 10 mL cold fixative solution (100% [v/v] cold methanol) was added to quickly halt transcription.
Warm the cold W5 solution to 23°C.
and quickly perfused transcardially with a cold fixative solution of 4% paraformaldehyde in 0.12 M sodium phosphate buffer (PB), pH 7.4, for 30 min. The brains were then removed from the skull and quickly sectioned in the coronal plane.
After we achieved temperature equilibrium, we poured into the solution 50 ml of 1°C cold normal saline or 50 ml of 1°C cold colloid solution in random order.
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