Forum - IACUC Approved Protocol

Here is the protocol! Keep in mind the surgery prep is not listed.

Imaging of neurons in the brain with a miniaturized fluorescent microscope. Animals will be anesthetized, and prepped for surgery as above. A craniotomy will be made over the region of interest and AAV-GCAMP6 virus injected. For deeper regions, the overlying cortical regions will be aspirated. The GRIN lens (which is the objective of the miniaturized microscope) (length 5 mm, diameter 1-2 mm) will be attached to magnetic baseplate (11 X 11 mm, < 0.3 g in weight), lowered into the desired region and dental cemented in place. The rest of the skull will be covered in dental cement. The animal will be allowed to recover. Two weeks later, the miniaturized microscope (<2g in weight) will be attached to the baseplate through magnetic contacts. The mouse will be released in the behavioral arena. The microscope will be connected with the computer using wires which will not impede the locomotion of the animal. The area imaged will be iluminated with a low-energy light emiting diode in the microscope and images saved to the computer. After the experiment, the microscope will be detached and the animal returned to its cage.

Thanks, Sergio