Clipping cartridges for CyoARM300

If there are no empty cartridges first unclip the used cartridges.

Note: All images are shown without liquid nitrogen in the workstation for easier visibility.

Figure 1: Scheme of the Cryo workstation for clipping grid

Figure 2: Closeup scheme of the magazine and vessel area

Figure 3: Closeup scheme of the specimen attachment area

Figure 4: Magazine vessel in cup.

You can find an empty magazine in the vessel wrapped in aluminium foil in the oven. Never touch it bare handed.

Figure 5: Picking up of the magazine

Figure 6: picked up magazine

You can pick up the magazine by putting a tweezer through the two front holes and grab it. (Figure 5 and 6)

Figure 7: Grabbing the magazine with its white pick up tool (topview)

Figure 8: Grabbing the magazine with its white pick up tool (sideview)

Now grab the magazine with the white pick up tool as shown in Figure 7 and 8

Figure 9: magazine being inserted into the workstation

Figure 10: Magazine properly aligned in the workstation

Put the magazine into the proper position in the workstation. If done well, the magazine is sliding smoothly along the rail (Figures 9 and 10).

Figure 11: Inserting the cup into the workstation

Grab the cup tightly with the carrying tool and insert it into the workstation. Use the pictogram as a guide to know the proper rotation. It should sit tight.
Cool down the workstation with liquid nitrogen. Once cold, make sure that the liquid nitrogen level is not higher than the specimen attachment area (Figure 1).

Figure 12: The white rotation tool is used to rotate the cup in the workstation

Use the white rotation tool (Figure 1) to turn the cup 90 degrees with the opening towards the magazine (Figure 12).

Figure 13: The white rotation tool is used to rotate the magazine toward the assembly site.

Figure 14: Magazine properly rotated

Figure 15: Safety pin of the magazine engaged

Use the white rotation tool to turn the magazine ~90 degrees with the opening towards the assembly site (Figure 13). Once in position (Figure 14), engage the safety pin (Figure 15)

Figure 16: Grid box, cartridge and C-ring inserted in the assembly station

Place the empty cartridge in the cartridge placement space (Figure 16).
Place your grid box in the specimen placement area (Figure 16).
Place the C-ring in the C-ring placement space (Figure 16).

Figure 17: Spreading of the white Specimen attachment tool using the rotation tool

Ensure that the tip of the specimen attachment tool is properly spread by inserting the rotation tool at the end (Figure 17).

Figure 18: Clipping tools and tweezer resting in the workstation.

Put a tweezer, the black C-ring adjustment tool and the white specimen attachment tool into the workstation and make sure they are cooled down before use (Figure 18).

Figure 19: Positioning of the cartridge on the specimen attachment area

Put a cartridge with the screw facing you and the square toward the left on the rail of the specimen attachment area (Figure 19). If properly positioned, the cartridge can slide easily sideward.

Figure 20: Positioning of the C-ring attachment plate

Align the screw of the cartridge, with the hole of the C-ring adjustment plate and put down the plate. Lock it with the rotation lock (Figure 20). Note: The rotation lock here is not essential but helpful

Figure 21: Preparation for C-ring clipping.

Insert one C-ring into the C-ring adjustment plate (Figure 21). Note: The C-ring does not need to be completely flat in position, but not totally tilted either.

Figure 22: Fitting of the C-ring in the adjustment plate.

Use the black C-ring adjustment tool to clamp the C-ring in the C-ring adjustment plate by pushing the tool straight down (Figure 22).

Figure 23: Control of C-ring preparation

Lift up the C-ring attachment plate and control that the C-ring is properly sitting in the plate (Figure 23). Important: It needs to be straight, otherwise clipping will fail.

Figure 24: Repositioning of the cartridge

Move the cartridge further left on the attachment position, so the open square is aligned with the square lock (Figure 24).

Figure 25: Locking of the cartridge

Lock the cartridge with the square lock (Figure 25). The cartridge should not be able to move sideward anymore.

Figure 26: Positioning of the grid on the cartridge

Put your cryo grid, top side upwards, onto the cartridge. Make sure its properly positioned in the groove (Figure 26)

Figure 27: Positioning of the C-ring attachment on top of the grid.

Put down the C-ring attachment plate with the C-ring, covering the cryo grid. It is vital to use the rotation lock to rigorously lock the attachment in position (Figure 27).

Figure 28: Clipping the C-ring onto the cartridge.

Use the white specimen attachment tool to clip the C-ring onto the cartridge, securing the cryo grid. Important: Come down straight with the tool, no twisting, no turning (Figure 28).

Figure 29: Control look of proper clipping.

Open the rotary lock and carefully lift up the attachment plate. If the Clipring is not completely on the cartridge you can put down the attachment plate again and use the white tool again for clipping. Otherwise you can lift up the plate completely. Important: Carefully check that the clipring is properly and straight on the cartridge. (Figure 29)

Figure 30: Clipped cartridge.

Open the square lock from the cartridge (Figure 30).

Figure 31: Cartridge tilting.

Slide the cartridge further left. It will automatically tilt toward you at the position shown in Figure 31.

Figure 32: Loosely positioned cartridge in the magazine.

Pick up the cartridge with a blunt tweezer and let it glide into the magazine (Figure 32). In the depicted image it is put in position 4.

Figure 33: Tight positioning of the cartridge into the magazine.

Use the straight egde of the white rotation tool to put the cartridge tightly into the magazine. The is a small resistance to overcome (Figure 33).
Repeat above steps for up to 4 samples.

Figure 34: Opening of the lock of the magazine

Open the lock of the magazine (Figure 34).

Figure 35: Moved down magazine.

Use the white rotation tool to turn the magazine down (Figure 35).

Figure 36: Magazine being slit into its vessel.

Use the flat side of the rotation tool to push the magazine into its vessel (Figure 36).

Figure 37: Rotating of the magazine vessel.

Once the magazine is in its vessel use the white rotation tool to lift up the vessel (Figure 37).

Figure 38: Rotated magazine vessel.

Once the magazine vessel is upright, fill it up with liquid nitrogen, so that the whole magazine is covered. No need to fill it up to the rim of the vessel (Figure 38) Important: Before doing the next steps, make sure the microscope is ready for grid insertion, i.e. it is not making images at the moment.

Figure 39: Position of the cryo cup.

Put the cryo cup next to the assembly workstation, have the pictogram on the lid oriented in the same direction as the magazine vessel; i.e. small and large circular structure (Figure 39)

Figure 40: Picking up of the magazine vessel.

Pick up the magazine vessel with the large grabbing tool (Figure 40). Important: Ensure a good grid to not drop the vessel.

Figure 41: Magazine vessel inserted into the cryo cup.

Position the magazine vessel in the cryo cup (Figure 41). Note the proper orientation with the pictogram.

Figure 42: Closed cryo cup.

Close the lid of the cryo cup and bring it to the microscope for insertion.
Clean up after finishing and put all the tools back into its box and drawer.