Protocol

1. Prepare a 1-liter bottle and a 500-mL bottle.
   
2. Place 100 mL/50 mL of 10x ACSF stock without glucose in the 1-liter/500-mL bottle.
   
3. Add 100 mL/50 mL of 10x bicarbonate, then add bottled DI water up to around 800 mL/300 mL.
   
4. Weigh and add the desired amount, as indicated below, of glucose and sucrose in each beaker. Use a small weigh boat and the more precise scale. To add the crystals to the solution in the beaker, add small amounts of bottled DI water into the weigh boat and carefully pour directly into the solution in the beaker without touching the rim. Wash the weigh boat for at least 3 times and pour out from different vertices of the weigh boat.
   
5. Add bottled DI water up to around 950 mL/450 mL and make sure everything is dissolved.
   
6. Use the osmometer and bottled DI water to achieve an osmolality in the range 295-300 mmol/kg. Add a clean stir bar and stir for 1 minute before taking the first measurement. For subsequent measurements, stir for at least 30 seconds after adding water. Take 2 stable readings for the final solution.
   
   

Regular ACSF

Low-glucose ACSF