Multi-session Awake Animal Imaging

Awake animal imaging is suited for multi-session imaging without the interference of anesthesia. Depending on the design of the experiment, reimaging can be obtained minutes, hours to days after the first view (see Note 13).

Carefully remove the silicone gel covering the skull and find the thinned region based on the brain vasculature map, and check the image quality with the TPLSM microscope. Skull re-thinning may be needed if the reimaging is done 3 days after the previous imaging.

Find the previously imaged region under the fluorescence microscope. Align the region according to the low magnification map under TPLSM, and then zoom in to higher magnification to further align the area. After the region is precisely aligned with the first view, take images with TPLSM.

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Yang G., Pan F., Chang P.C., Gooden F, & Gan W.B. (2013). Transcranial Two-Photon Imaging of Synaptic Structures in the Cortex of Awake Head-Restrained Mice. Methods in molecular biology (Clifton, N.J.), 1010, 35-43.

Publication 2013

Anesthesia Animal Brain map Fluorescence microscope Microscope Silicone gel Skull

Corresponding Organization :

Other organizations : New York University

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Variable analysis

independent variables

Time between first view and reimaging (minutes, hours, days)

dependent variables

Image quality

control variables

Skull condition (whether re-thinning is needed)

controls

Positive control: Checking image quality with TPLSM microscope
Negative control: Not mentioned

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