0.9 numerical aperture water immersion objective

Slices were placed in a submersion chamber on an upright microscope, viewed with an Olympus 60× (0.9 numerical aperture) water immersion objective with DIC and infrared optics, and perfused with ACSF at a rate of ~7ml/min. Drugs were dissolved in ACSF and applied by bath perfusion. Patch pipettes were constructed from thick-walled borosilicate glass capillaries and filled with an internal solution containing (in mM): CsCl 130, NaCl 4, CaCl₂ 0.5, HEPES 10, EGTA 5, MgATP 4, Na₂GTP 0.5, QX-314 5, pH adjusted to 7.2 with CsOH. Electrode resistance was 4 to 10 MΩ. Cells were rejected if access resistance was greater than 15 MΩ.

Slices were placed in a submersion chamber on an upright microscope, viewed with an Olympus 60× (0.9 numerical aperture) water immersion objective with DIC and infrared optics, and perfused with ACSF at a rate of ∼7ml/min. Drugs were dissolved in ACSF and applied by bath perfusion. Patch pipettes were constructed from thick-walled borosilicate glass capillaries and filled with an internal solution containing (in mM): CsCl 130, NaCl 4, CaCl₂ 0.5, HEPES 10, EGTA 5, MgATP 4, Na₂GTP 0.5, QX-314 5, pH adjusted to 7.2 with CsOH, E_Cl- = ∼ 0mV. Electrode resistance was 4 to 10 MΩ. Cells were rejected if access resistance was greater than 10 MΩ, or if the access resistance changed by more than 20% across a given experiment. Details for the number of cells and the number of animals used for various electrophysiological experiments are supplied in Table 2.