Pulling micropipettes

[NOTE: Important preliminary puller information]

Right Puller: This machine needs a new memory chip (as of 08/22/2022) and is much slower than the one on the left. When using it, it doesn't necessarily matter which program you use as all of the parameters will need to be typed in manually anyways.

After typing in the program number, you need to clear all of the data values prior to plugging in new ones. You can do this by clicking on the CLR button, and then clicking 1 for yes. Then use the NEXT button to click through the eight different lines and make sure that no numbers are left. If the numbers are still there, you may need to pick a different program number and repeat this cycle again. If the numbers are gone, then you're ready to set up the parameters for the pipette.

Left Puller: This machine is faster than the right one and has a functional memory card. The program number you select should be consistent with the parameters of the pipette you will be pulling, so reprogramming it every time isn't necessary. You should always check to make sure they're correct though. If the numbers don't line up with what you want, plug in the correct ones.

Pulling injection pipettes

Get the injection capsule tubes from the "Manuals/Capillary Tubes" drawer (what they look like).
Select a capsule tube.
Open the hood of the puller.
Using a Kimwipe, wipe down the capsule, primarily the area that will become the pipette tip (the middle).
Carefully slide the capsule tube into the slot indentation and delicately feed it through the heating box, into the other side. (Capsule in slot), (capsule outside heat box), (capsule passing through heat box).
Using your fingers, hold the capsule tube in place while tightening the clamps on either side. Don't tighten them fully, just enough so that the tube is held in place (looks like this).
Maneuver the tube so that it covers equal distance on both sides. The ends should reach the same area, using the bolt above for reference.
You now need to unlock the pulling mechanisms. These are the small metal tabs on the far left and right (here). To unlock: push one tab in, slide the pulley apparatus towards the heating box, and release the tab. Repeat for the other side. (Before unlocking), (after unlocking).
Now you're ready to fully clamp the tube. Using two fingers, push the metal lips (below the pulleys) towards the heating box, so that the two pulleys are pressed against heating box. Use the "(after unlocking)" link above for reference. Keeping this position, fully tighten the clamps so that the capsule tube is unable to move.
If you let go of the metal lips of the pulleys, and they don't move away from the heating box, then you've done it correctly.
Shut the lid of the puller and turn the power button on. The button's on the left side of the machine.
Using the keypad, type in your program number. If using the left rig, use 1. If using the right rig, use 2 or 5 (although it doesn't matter, these programs tend to work easier when plugging in parameters).
Click ENTR

If using the left machine, double check that the parameters for heat, velocity, time, pull, and pressure are what you need them to be. To check/change pressure's value, click CLR, then 0, and then 2. Type in the correct values if any of them are wrong. Click pull when ready.
- The left rig should finish the pipettes relatively quickly. You'll see light emitting from inside the heat box and will hear a pop once the pull occurs. Don't open the puller until you're sure that the pipettes have been pulled.
If using the right machine, click CLR first, and then 1. This clears out the previous parameter values. Use the keypad to type in the desired values for heat, velocity, time, pull, and pressure. To check / change pressure, click on CLR, 0, and then 2 in succession. Hit ENTR after typing in the pressure. Using the NXT and LAST buttons, check that only the first line has numbers plugged in. Click pull when ready.
- The right rig will take several minutes to pull, but it too will emit light and make a popping noise. Give it plenty of time to finish and don't open it before it's done.

You should see two newly formed pipettes in the clamps if the pulling was successful. Free them by unscrewing the clamp while pinching the pipette. It should come free.
Examine them under the MF-830 microscope to check for quality. They should look like fine tipped points.

Pulling TEVC pipettes (channel 1 / channel 2)

Get the TEVC capsules from the "Manuals/Capillary Tubes" drawer. (They look like this).
Select a capsule tube.
Open the hood of the puller.
Using a Kimwipe, wipe down the capsule, primarily the area that will become the pipette tip (the middle).
Carefully slide the capsule tube into the slot indentation and delicately feed it through the heating box, into the other side. These are shorter than those used for injections, so it may be a little harder to line it up evenly on either side.

NOTE: Much of this process is homologous to pulling injection pipettes. Looking at pictures from that protocol may help you determine if you're doing this correctly. Just keep in mind that the pipettes will look different.

Using your fingers, hold the capsule tube in place while tightening the clamps on either side. Don't tighten them fully, just enough so that the tube is held in place.
Maneuver the tube so that it covers equal distance on both sides.
You now need to unlock the pulling mechanisms. These are the small metal tabs on the far left and right (here). To unlock: push one tab in, slide the pulley apparatus towards the heating box, and release the tab. Repeat for the other side. (Before unlocking), (after unlocking).
Now you're ready to fully clamp the tube. Using two fingers, push the metal lips (below the pulleys) towards the heating box, so that the two pulleys are pressed against heating box. Keeping this position, fully tighten the clamps so that the capsule tube is unable to move.
When you let go of the metal lips of the pulleys and they don't move, then you've done it correctly.
Shut the lid of the puller and turn the power button on. The button's on the left side of the machine.
Make note of which puller you're using [Left or Right]. Read through the preliminary instructions at the top of this page for clarification on how to program your chosen puller.
After programming, click pull.
After it's finished running, open the hood and you should see two, newly formed, fine tipped pipettes.
Free them by unscrewing the clamps and pulling them free.
Examine them under the MF-830 microscope to check for quality. They should look like this.

DO NOT BREAK the channel 1 or channel 2 pipettes! They serve a different purpose than the injection pipettes.

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