Secure a mouse in a stereotactic frame with a surgically exposed brain containing a pre-implanted glioblastoma spheroid.
Apply saline to the exposed brain to maintain tissue hydration.
Position the flexible probe over the saline layer, ensuring its contact pads rest on the mouse's back and the electrodes align above the glioblastoma.
The probe consists of a thin, flexible polymer substrate with patterned gold electrodes coated with biocompatible conductive polymer, ensuring optimal electrical performance and tissue compatibility.
Absorb excess saline, leaving only a thin layer.
Apply silicone adhesive to stabilize the electrodes, then place a cover glass.
Apply dental cement around the cover glass and secure the contact pads to the mouse's back.
Construct a cement well for fluid insertion.
Later, administer post-surgical analgesia and maintain thermal support for the mouse's recovery.
The implanted electrodes deliver electrical pulses above the glioblastoma for targeted pulsed electric field therapy in neurotherapeutic applications.
Place a drop of DPBS to cover the craniotomy. Place the flexible electrode onto the drop of DPBS and gently place the back of the probe with the contact pads onto the mouse's back. Absorb the DPBS drop with a small piece of paper until the probe can lay flat on the dura and follow the curvature of the brain, ensuring the small layer of saline remains below the electrodes to act as a barrier against glue spillover.
Place a small drop of silicone adhesive onto the probe and cover it with a five millimeter round cover glass. Push the cover glass down until the silicone is evenly distributed and the distance between the cover glass and the probe is minimal. Then wait 30 seconds for the silicone to solidify.
To secure the cover glass, quickly apply superglue on its sides and push it down until the glue becomes solid. Apply super glue at the probe's neck using a toothpick taking care that the super glue gets drawn under the neck to provide stable support. Cover the skull with dental cement to build a chronic cap and take special care to cover the edges of the cover glass only.
Lift the back of the probe and apply cement underneath the neck of the probe. Rest of the probe onto the cement before it cures. Push down the neck of the probe gently to place its surface at the same level as the cover glass and not in the way of the microscope objective during the experiment.
Cover the top of the probe neck with not more than 1.5 millimeters of dental cement layer to achieve a firm hold on the probe. Build a cement well presenting a 1.5 millimeter ridge at one to two millimeters around the cover glass to create a basin for the immersion fluid for the two photon imaging. After the cement has cured, administer postsurgical analgesia and keep the animal warm until recovery by wrapping it in a paper towel and placing it close to an infrared light bulb.
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