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The research using new born rat brains was conducted according to national guidelines under the UK, Animals (Scientific Procedures) Act 1986 and all animals were born in the University of Leicester Biomedical Sciences Unit.
An in vitro BBB model using primary cells isolated from rat brains was established.
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The infant rat brains were removed for analysis 6 weeks after the hypoxic ischemic insult.
The rat brains were stored in 20% sucrose at 4°C until freezing, and frozen brains were cut on a microtome into 40-μm sections in series of six.
Rat brains were fixed by intracardial perfusion with 4% paraformaldehyde for 10 min. The brains were then removed and post-fixed in 4% paraformaldehyde overnight.
Rat brains were harvested for 2,3,5-triphenyltetrazolium chloride (TTC) staining analysis to evaluate infarction volume at 14 days after MCAO (BMSC group, n = 15; vehicle group, n = 15).
Rat brains were embedded in resin and sectioned serially to allow quantitative 3-D analyses of single neurons or groups of neurons.
In control experiments, rat brains were injected with PKH 26-labeled UCM cells that had been lysed by repeated sonic disruption.
PET data in rat brains were mostly consistent with LC-MS/MS findings, and rat and monkey PET scans demonstrated that [11C]TASP0410457 was superior to [11C]TASP0434988 for high-contrast H3R PET imaging.
For in situ hybridization, rat brains were dissected and rapidly frozen.
Membrane fractions from adult rat brains were isolated by sequential centrifugation steps.
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