III. Recording CA1 neurons in organotypic slices

Procedure

1. Refer the lab protocol to make organotypic slices.

2. For thick slices (350 ~ 400 um for LTP protocol), don’t put them in 37 degree incubator for initial incubation.

According to lab protocol, 37 degree is only for 250 um slices (Standard slice). SCJ ver. All were incubated in the 35 degree incubator

3. SCJ Ver. Use only DIV 6-7 slices for consistent results. DIV 8 neurons would make your error bars bigger.

4. Div 6-7 neurons have -55 ~ -65 mV holding potentials right after breaking.

5. Whole cell cap is usually 10-20 pF.

6. Cell body size at same days (Div 6-7 organotypic : P14 acute)

Organotypic slice > acute slice

7. Membrane weakness (Div 6-7 organotypic : P14 acute)

Organotypic slice < acute slice

8 Alive cell numbers (Div 6-7 organotypic : P14 acute)

Organotypic slice >>>>>>>>>>>>> acute slice

9. Physiological condition (Div 6-7 organotypic : P14 acute)

Organotypic slice <<<<<<<<<<<< acute slice

10. For recording EPSCs, stimulating electrode should be located at least 100 um away from cell bodies.

11. The stimulating intensity should be adjusted from lower current intensity.

12. A tissue holder in the recording chamber can damage Schaffer collateral pathways. Be careful when you hold slices with holder.

SCJ ver. Regular external recording solution (organotypic & acute slices)

pH 7.3~7.4

• To reduce recurrent in the organotypic slice recording, use 5uM 2-chloroadenosine.

• To block GABA receptors, 5~10uM Bicucu. can be used in acute slices and 2~5 uM bicucu. in organotypic slices