Internal Saline
| Reagent | Final concentration |
|---|---|
| K-aspartate | 125 mm |
| CaCl2 | 0.1 mm |
| EGTA | 1.1 mm |
| HEPES | 10 mm |
| MgATP | 4 mm |
| Na3GTP | 0.5 mm |
| Biocytin hydrazide (optional to dye-fill neurons) |
13 mm |
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1. Adjust the osmolarity of the solution to 265 mOsm by changing K-aspartate concentration (the lower osmolarity of the internal saline relative to the external facilitates patch rupture). If you do not intend to dye-fill, replace biocytin hydrazide with an equal molar quantity of K-aspartate.
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2. Adjust the pH to 7.3 and filter through a 0.2-µm membrane to remove any particulates.
- © 2013 Cold Spring Harbor Laboratory Press
