Tzahor Lab


Needle preparation:

    1. Use kwik-fil capillaries MTW100-4 by World Precision Instruments, Inc.
    2. Take Petri dish with plasticine.
    3. Place the capillary in the machine and choose program 70.
    4. Close the top and press Pull. Wait until it goes back to the initial message screen.
    5. Take the needle out and put it on the plasticine in the dish. You can start over with new capillary

Loading the needle:

  1. Prepare your plasmid in the right ratio (centrifuge it first).
  2. Add a tiny amount (only with the tip of the tip) of fast green.
  3. Cut the top of a long 20μl eppendorf tip and connect it to a syringe.
  4. Upload the plasmid and inject it to a needle.
  5. Break the needle under the binocular using tweezers or tungsten knife.


  1. Connect positive charged red wire (brown) to the upper electrode.
  2. Connect negative charged black wire (green) to the bottom electrode (round one). Use masking tape and connect it to the binocular so it will not move.
  3. Adjust the micromanipulator (the good one…).
  4. Fill the bottom electrode with Hank's solution all the way to the top.
  5. Bring the upper electrode into the Hank's solution only to the point it is covered with solution.
  6. Turn on the elctro-porator and test it: 30V, 25ms, 2 pulses and see if u can see small bubbles (you should…). If you don't see them check your connection and see if there is a big bubble stuck in the bottom electrode.
  7. Set the configurations: • St.2-3 – 6V, 25ms, 2 pulses (500μs interval) • St.6-8 - 7V-8V, 25ms, 3 pulses (500μs interval)
  8. Soak the embryo gently (from new-culture) in the Hank's solution in the bottom electrode. Clean it (very jently) if needed.
  9. Adjust the embryo so it is beside the bottom electrode – you need to see the embryo clearly and to be able to inject comfortably.
  10. Disassemble the syringe and draw out the piston (always do this when the syringe is disassembled!).
  11. Connect the syringe, insert the needle to the bottom, inject to the desired position and elevate it slowly until you see green circle accumulate.
  12. Place the embryo just above the bottom electrode and the upper one precisely above it (see step 5).
  13. Press Start.
  14. Every few embryos check if you can see the bubbles. When the Hank's becomes pink replace it.