Installation of Optical Windows in the Brain of Transgenic Mice to Image Neural Activities

Begin with an anesthetized, genetically altered mouse with its head secured in a head-holding adaptor.

The mouse brain expresses a fluorescent protein that enables the detection of neural activities.

Shave and sequentially sterilize the scalp before cutting it open.

Remove connective tissue to expose the skull.

Mark the target areas and apply a layer of glue.

Use a microdrill to thin the skull on the marked area.

Remove the piece of the skull to expose the brain.

Apply artificial cerebrospinal fluid to keep the brain moist.

Install an optical window in the area and seal it with glue, which provides a base for the cement attachment.

Apply black dental cement to block the light.

Repeat the procedure on the other side of the skull.

Allow the animal to recover.

To image neural activity, place the secured anesthetized mouse under the microscope and acquire images.

Confirm a lack of response to toe pinch on an anesthetized 20 to 25 gram, 1 and 1/2 to 2 and 1/2-month-old Thy1-GCaMP6s mouse and apply ointment to the animal's eyes. Place the mouse on a heating pad covered with a sterile drape, and use a head-holding adapter for mice to stabilize the head. Shave most of the scalp with a doub