Biology Reference
In-Depth Information
2.1. Materials
and Instruments
Vibratome equipped with a sapphire knife (Leica 1200, Leica
Microsystem, Richmond Hill, Ontario, Canada).
Upright light microscope with a fi xed stage.
Computer
Amplifi er (Multiclamp 700 A, Molecular Devices, Foster City, CA,
USA)
Digitizer (Digidata, 1320, Molecular Devices)
Software Clampex 8 for data acquisition (Molecular Devices).
Software Clampfi t 9.2 for data analysis (Molecular Devices).
Bipolar concentric tungsten electrode for electrical stimulation
(FHC, Bowdoinham, ME, USA).
A digital stimulator controlled by a computer program (STG 1001,
Multichannel System, Reutlingen, Germany).
Patch glass capillary tubing (Warner Instrument Corp.).
Flaming/Brown micropipette puller (Sutter Instrument Company,
USA)
Peristaltic pump with tubings for drainage (Easy-load Master fl ex
pump, Cole-Palmer Inst. Co. USA)
Plastic tubings of various diameters.
Artifi cial cerebrospinal fl uid (CSF) with the composition (in mM):
124 NaCl, 5 KCl, 1.25 NaH
2
PO
4
, 2.5 CaCl
2
, 1 MgSO
4
, 26
NaHCO
3
, and 10
D
-glucose.
Compressed gas tank with medical grade 5% CO
2
balanced O
2
.
Tissue chamber
Razor blades
2.2. Slice Preparation
Anesthetize the rat with ketamine/xylazine (90/10 mg/kg
weight), decapitate the animal with a small animal guillotine,
and quickly remove the brain and place it into ice-cold artifi cial
CSF oxygenated with 95% O
2
and 5% CO
2
.
Cut the cerebellum and anterior frontal lobe with a razor blade
without damaging the hippocampal region of the brain.
Glue the brain block onto the holder, and put it into the slicing cham-
ber of vibratome fi lled with ice-cold oxygenated artifi cial CSF.
Divide the brain into two halves by cutting through the midline
with a blade.
Cut 400
m thick transverse sections through hippocampus using
vibratome and transfer them to tissue chamber with oxygenated
artifi cial CSF at room temperature.
Keep the slices at room temperature for 1 h before recording.
μ
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