Fifteen to 43 days following ChR2 virus injection, brain slices containing mPFC and/or NAcc were prepared as previously described36 (link). Briefly, animals were decapitated under isoflurane anesthesia and brains rapidly removed and immersed in ice cold cutting solution perfused with 95% oxygen-5% carbon dioxide. Two-hundred μm-thick coronal sections containing mPFC and/or NAcc were then cut using a VTS-1000 vibrating blade microtome (Leica Microsystems). Slices were kept in oxygenated cutting solution at 32° C for 1 hour before being transferred to a recording chamber with regular artificial cerebrospinal fluid (aCSF). Slices were imaged using a Leica DM-LFS microscope (Leica Microsystems) captured with SimplePCI software (Hamamatsu Corp.) for areas of strong fluorescence within the target region of interest (mPFC or NAcc), and recordings performed as follows:
Protocol detail
Visualizing ChR2 Expression in mPFC and NAcc
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Anesthesia
Animals
Brains
Carbon dioxide
Cerebrospinal fluid
Cold
Fluorescence
Isoflurane
Microscope
Microtome
Oxygen
Virus
Corresponding Organization :
Other organizations : Emory University
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Variable analysis
independent variables
- Time (15 to 43 days) following ChR2 virus injection
- Neuronal activity in mPFC and/or NAcc regions
- Cutting solution (oxygenated)
- Recording chamber (with regular artificial cerebrospinal fluid, aCSF)
- Slice thickness (200 μm)
- Slice incubation temperature (32° C)
- Positive control: Not explicitly mentioned.
- Negative control: Not explicitly mentioned.
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