This article details a method for the targeted delivery of therapeutic agents into the rodent brain using microinjection techniques. The procedure emphasizes minimizing stress in the animal and ensuring sterility throughout the process.
Begin with a sterile water-filled microinjection setup consisting of a syringe connected to a microinjector via plastic tubing.
Retract the syringe plunger slightly to create an air bubble in the tubing.
Submerge the microinjector tip into the therapeutic agent and pull the plunger to aspirate the solution.
Take an awake rat with a protective head cap and two pre-implanted guide cannulae inserted into the specific target regions of the brain.
Hold the rat's chest against the experimenter's chest to minimize stress.
Clean the area around the cannulae with a sterile solution.
Insert the drug-loaded microinjectors into the guide cannulae to reach the target sites.
Start the drug infusion and monitor its delivery by tracking bubble movement.
Once the injection is complete, hold briefly, then withdraw the microinjectors. Insert the obturators into the cannulae to prevent contamination.
This method enables the targeted delivery of therapeutic agents into the rodent brain.
Pull the syringe plunger back 0.2 microliters to create a bubble between the sterile water and the PE 20 tubing and the solution to be injected.
After that, place the micro injector into the reagent to be injected and slowly pull the syringe plunger to the backstop. To prepare the animal for injection, hold the rat's chest against the experimenter's chest. Clean the area around the cannula with a cotton applicator soaked in Betadine.
Swab the head cap three times with Betadine and then swab with ethanol two times. Then place the filled micro injector into the cannula. Press start on the micro injection pump and monitor the bubble movement.
When the injection is complete, remove the micro injector from the cannula and replace with obturator.
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