Surgery¶

The type of surgery required to prepare for fUSI recordings will vary depending on your recording modalities. From simple shaving/depilation of the imaged area for anaesthetized cases, to skull removal and replacement with an imaging window, through skull thinning or headpost fixation, surgeries of various difficulties will be required depending on your experimental design.

Here we list different preparations ordered by increasing invasiveness:

• Low:
  • Anesthesia + shaving - in mice only
    • The animal is first anaesthetized and shaved (with scissors first, depilation cream then)
    • The animal is placed on the stereotaxic frame
    • Gel is poured on the skin and images can be acquired
• Intermediate:
  • Anesthesia + skull thinning - in rats
    • The animal is anaesthetized and put on the stereotaxic frame
    • The skin on top of the skull is removed and the external part of the skull is thinned out with a surgical drill
    • Gel is poured on the thinned skull window and images can be acquired (it is recommended to wait at least for one day for recovery before imaging the animal)
  • Awake transcranial imaging, with headpost implant - in mice
    • The animal is anaesthetized and put on the stereotaxic frame
    • The skin on top of the skull is removed
    • [optional] Some holes are drilled superficially to allow fixation reinforcement by screws
    • The headpost is placed and the cemented in place
    • [optional] Some cyanoacrylate glue is poured on the skull to protect it
• High:
  • Awake imaging through chronic craniotomy
    • The animal is anaesthetized and put on the stereotaxic frame
    • The skull is removed while keeping the dura mater intact
    • The implant is put on top of the brain and cement used to seal the implant on the head
    • A protective layer of of removable material (silicon) within the imaging chamber is used to protect the implant between two imaging sessions

Protocols¶

Detailed protocol for imaging in awake head-fixed mice, including details of the cranial window implantation.

Whole-brain functional ultrasound imaging in awake head-fixed mice | Nature Protocols

An open-source 3D-printable cranial implant to do both fUSI and two-photon imaging through the same window

The COMBO window: A chronic cranial implant for multiscale circuit interrogation in mice | PLOS Biology