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RWD Protocol of Different Methods for Intracranial Injection

RWD Protocol of Different Methods for Intracranial Injection
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RWD Protocol of Different Methods for Intracranial Injection - 1

Protocol of Different Methods for Intracranial Injection In the field of basic brain science research, intracranial stereotaxic injection is the most direct way to achieve precise injection in target brain regions, and has become an important part of most animal experiments such as injection of viruses, cells, protein molecules, drugs, labeled dye probes, etc. Combined with the matching optical fiber, it also enables optical stimulation or neuronal signal recording of the target brain region during, or after the injection. This method is widely adopted in animal models of human neurological diseases, advanced brain functions, emotions, cognition and other related studies. Intracranial stereotaxic injection is usually applied in single administration and multiple administrations. During the experiment, the appropriate method can be selected according to the frequency of injection, the time of onset and the duration of action. In this paper, we will elaborate on these two methods in details, including the SOP and tips. SOP of Cranial Single Administration 1 Instruments Stereotaxic Apparatus Laboratory Animal Anesthesia machine Micro syringe pump -1-

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RWD Protocol of Different Methods for Intracranial Injection - 2

Micropipette puller 2 Process and procedure 2.1 Pre-surgery animal anesthesia Turn on the animal anesthesia machine. Place the rat in the induction box to conduct induced anesthesia. Once the rat is under anesthesia, take it out from the induction box and transfer to the anesthesia mask at the stereotaxic instrument with a heating pad. Apply[XY1] eye ointment to their eyes and prepare the skin. 2.2 Fix the skull 2.2.1 Put the rat’s incisor into the hole of the mask and gently screw the nose clip. 2.2.2 Move the ear bar on one side to the appropriate position and screw it into place. Hold...

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RWD Protocol of Different Methods for Intracranial Injection - 3

2.4.3 Loosen the chuck of the micropipette injection pump to gently push the glass capillary to the top, and then tighten the chuck. 2.4.4 Set parameters for emptying speed and emptying volume in the control unit and click to execute emptying. 2.4.5 Immerse the capillary tip in the solution to be injected. Set parameters for filling speed and filling volume in the control unit and click to perform filling. 2.4.6 Fix the micropipette injection pump on the operating arm. After resetting the capillary tip to zero, move the operating arm above the target site and slowly descend to the target...

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RWD Protocol of Different Methods for Intracranial Injection - 4

SOP of Cranial Multiple Administration 1 Instruments Stereotaxic Apparatus Drug delivery cannula Micro syringe pump 2 Process and procedure 2.1 Pre-surgery animal anesthesia 2.1.1 Turn on the animal anesthesia machine. Place the rat in the induction box to conduct induced anesthesia. Once the rat is under anesthesia, take it out from the induction box and transfer it to the anesthesia mask at the stereotaxic instrument with a heating pad. Apply eye ointment to their eyes and shave the skin on their head for preparation. 2.2 Fix the skull 2.2.1 Put the rat’s incisor into the hole of the mask...

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RWD Protocol of Different Methods for Intracranial Injection - 5

micro drill to touch the Lambda to record the value of DV. If the absolute value is less than 0.03mm, the skull can be regarded in the horizontal position; if greater than 0.03mm, the height of the adapter needs to be adjusted, and the positions of Bregma and Lambda need to be re-calibrated until the absolute value of Lambda DV is less than 0.03mm. 2.3.2 Move the cranial drill to the middle point of the Bregma and Lambda line and expand the left and right sides by 0.3mm. Touch the cranial surface gently, and record the craniofacial DV values at the left and right points respectively. If the...

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RWD Protocol of Different Methods for Intracranial Injection - 6

3 Tips 3.1 When drilling holes at the position where cranial screw are embedded, special attention shall be paid in order not to drill through the skull and avoid brain tissue damage. Besides, the drilling location should be kept as far away as possible from the implanting area of the cannula to avoid the cranial screw blocking. 3.2 For bilateral implantation, implant the right single cannula first to prevent the spatial obstruction. 3.3 Apply the dental cement layer by layer to solidify the implantation.

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