Intracellular Recording In Vivo and Patch-Clamp Recording on Brain Slices - Animal Models of Acute Neurological Injuries: Injury and Mechanistic Assessments - page 110

Fig. 7. An example of brain slice recording setup.

Fig. 8. Schematic drawing of a device for securing slice in recording chamber.

2.3. Procedures

1. Several solutions, such as the sucrose solution, ACSF, and
intracellular solutions, are prepared at the beginning of the
experiments. All solutions should be prepared with double-
distilled water (>18 MW cm). In addition, CaCl 2 should be
added at the last step to prevent precipitation. Both sucrose
solution and ACSF are prepared fresh daily. The intracellular
solutions can be stored in aliquots (0.5-1.0 ml) at −20°C, up
to 1 month. The sucrose solution is used for transcardiac per-
fusion and for cutting slices. This solution contains (in mM)
230 sucrose, 26 NaHCO 3 , 2.5 KCl, 1.25 NaH 2 PO 4 , 0.5 CaCl 2 ,
10 MgSO 4 , and 10 glucose, pH 7.4, and 290-305 mOsm/L,
oxygenated with 95% O 2 and 5% CO 2 . The ACSF is used for
incubating slices and used as normal extracellular solution during
recordings, which contains (in mM) 130 NaCl, 3 KCl, 2 CaCl 2 ,
2 MgCl 2 , 1.25 NaH 2 PO 4 , 26 NaHCO 3 , and 10 glucose, pH

2.3.1. Brain Slice
Preparation