aCSF solution

Reagent	Final concentration	Amount
10× aCSF	1×	100 mL
1 M CaCl2 solution	2 mM	2 mL
1 M MgCl2 solution	1 mM	1 mL
D-(+)-Glucose	20 mM	3.6 g
ddH2O	N/A	Fill up to 1,000 mL
Total	N/A	1,000 mL

Add 1 M CaCl₂ solution and 1 M MgCl₂ solution after diluting 10× aCSF solution with ddH₂O and bubbled with a mixture of 95% O₂ and 5% CO₂ for 10–30 min at 20°C–25°C.

Although the solution may be stored at 4°C for one week, we recommend to prepare the fresh solution on the day of use.

Osmolarity of the solution is 310 ± 5 mOsm.

pH of the solution falls to 7.2–7.3 after bubbling with a mixture of 95% O₂ and 5% CO₂ for 10 min at 20°C–25°C .