1/2 uL Syringe & Glass Micropipette Priming Protocol
COLLECT:
Infusion tubing cut into 7 in. segments
22 gauge metal tubing cu into ~2.5 in. segments use the dremel down to smooth and widely open ends
23 gauge needle (green)
30 gauge needle (pale pink)
1 mL syringe
Mineral oil placed in small weigh boat
Kim wipes
.5uL syringe & glass micropipette adapter
Pre-opened micropipette (puller in Rebec lab HEATER: 6-7 MAGNET: 5)
PROCEDURE:
Once you have all items collected make sure you clear off some space for you to work. Begin by filling up the 1mL syringe with mineral oil while making sure all bubbles are omitted within the syringe. Attach the 23-gauge needle onto the syringe and draw up the mineral oil until it dispenses out of the 23g needle. At this point you should have a full 1mL syringe with a 23g needle attached full of mineral oil without any air bubbles. Next, insert ~ 1/6 inch of a single segment of 22 gauge metal tubing into one of the ends of the infusion tube segment. After this is done, completely insert the metal part of the 23-gauge needle into the available end of the infusion tubing. Carefully and slowly press on the syringe and draw up the mineral oil until you see it appear on the open end of the 22g metal tube. Now insert the 22g metal tube into the available end of the .5uL syringe adapter and slowly begin the fill the inside compartment of the adapter with mineral oil. You should begin to feel a force pushing out the 22g metal tubing at which point you should begin the withdraw the 22gh metal tubing while still dispensing mineral oil. In the end you should see the inside of the adapter full of mineral oil with NO air bubbles. Now it is time to prime the glass micropipette needle. Detach the 23-gauge needle from the 1mL syringe and attach a 30-gauge needle. Make sure there are no air bubbles when dispensing from the 30-gauge needle. Insert the 30g needle into the wide end of a previously prepared glass micropipette. Begin filling the micropipette with mineral oil making sure it fills the pointy tip all the way up. Once it has carefully begun the extract the 30g needle while still slowly and carefully dispensing mineral oil. In the end you should have a full glass micropipette with no air bubbles.
Once ready to inject carefully unscrew the top nut of the adapter and also detach the compression accessory from the adapter. You should see a small holder for your glass micropipette needle. Carefully place the wide end of the glass micropipette into the holder. VERY carefully place the compression accessory around the glass micropipette making sure to have the narrower end of the compression accessory facing down. Push it down into place. Lastly carefully place the top nut of the adapter around your glass micropipette needle and screw it down tightly. You now have a fully primed and ready to use .5uL glass micropipette needle and syringe.
WATCH VIDEO OF PROCEDURE HERE:
https://www.youtube.com/watch?v=QYUnf1ETVKk&feature=youtu.be