Microinjection pulling

Chen, J., Marmur, R., Pulles, A., Paredes, W., and Gardner, E.L., Ventral tegmental microinjection of A9-tetrahydrocannabinol enhances ventral tegmental somatodendritic dopamine levels but not forebrain dopamine levels evidence for local neural action by marijuana s psychoactive ingredient, Brain Res., 621, 65, 1993. 

Adjust the value of the current through the heating filament to your individual needs (usually this value should not vary more than 10% from the value recommended by the supplier) and pull the micropipette. The tip diameter of the micropipette decreases with increasing current and vice versa. Capillaries with an outer tip diameter between 0.2 and 1.0 //m are easily available with the puller used. Two micropipettes will be obtained after 20-30 sec. Both can be used for microinjection because they are identical with respect to their physical parameters. 

Place the depression slide in the incubator for 5 min. During this incubation, pull a microinjection needle on the pipette puller, and place the base in DNA solution for filling. After the tip is filled, fill the shaft behind the DNA with Fluorinert using a spinal needle. 

Prepare dissection needles. Wear gloves to avoid contaminating the glass needles. Pull each capillary tube into two needles using any program for microinjection needles on a needle puller (the ends are broken off later, so the exact profile of the needle is not critical). 

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