Table 2 Critical steps in the brain slice experiment
From: The brain slice method for studying drug distribution in the CNS
Experimental stages | Critical steps |
|---|---|
Preparatory steps | Control the pH, osmolarity and oxygenation of the aECF |
Take into account the pKa values of the compounds when selecting the drugs to be investigated in one cassette | |
Do not take longer than 1 minute to extract the brain | |
Keep cold-chain during the brain slicing procedure | |
Preserve the brain slices in ice-cold oxygenated aECF before starting the incubation | |
Incubation | Keep oxygenation, temperature and stirring constant during the incubation |
Preparation of the samples for bioanalysis | Make sure all minor debris from the brain slices has sedimented before taking the aECF samples after the 5 h incubation |