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The slices are moved, using a micro spatula, to the brain slice storage beaker filled with oxygenated blank aECF which is kept in an ice bucket before the incubation.
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For brain slice preparation and storage, a low-sodium oxygenated solution containing (in mM) 64 NaCl, 120 sucrose, 25 NaHCO3, 10 glucose, 2.5 KCl, 1.25 NaH2PO4, 0.5 Cand2 and 7 MgCl2 was used.
Use a paring knife to cut the fruit into equal-sized slices for easier storage.
The incubation-equilibration process is started by gently transferring the 6 (rat) or 10 (mouse) brain slices from the storage beaker (using a micro double-ended spatula) into one 45 mm high, 80 mm diameter, flat-bottomed glass beaker (Duran Group, VWR, Sweden) containing 15 ml (rat) or 10 ml (mouse) of the aECF containing the selection of drugs to be investigated.
Freeze slices if longer storage required.
However, the preparation and storage of slices, as well as the flow rate of the solution during recordings, were altered: slices were kept in an interface-type chamber before recordings, and a higher flow rate was used in the recording chamber.
Slices were placed in a storage chamber containing continuously oxygenated slicing solution at 35°C, which was allowed to come to room temperature naturally.
In the case of Bafilomycin incubation, slices were pretreated in storage ACSF containing 45 μ m Bafilomycin A1 for 1 2 h prior to recording.
However, after 20 days of storage, the slices that underwent blanching and oven drying retained 83% of the initial amount of β-carotene, compared to only trace amounts found in the samples that were not blanched prior to the drying process, suggesting that blanching may improve carotenoid stability over time.
Put the plum slices in a freezer storage bag and freeze.
Ring-fencing of each slice operational resources (e.g., storage, processor, operational memory), so that one slice cannot exhaust other slice's resources in any situation.
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