Cellink BIO X User manual

Boston, U.S.
100 Franklin St.，
Boston, MA 02110
Gothenburg, Sweden
Arvid Wallgrens Backe 20,
Gothenburg, 41346
Virginia, U.S.
2000 Kraft Dr., Suite 2125
Blacksburg, VA 24060
Kyoto, Japan
46-29 Yoshida-Shimo Adachi-
cho, Sakyo-ku, Kyoto
www.cellink.com | +1 (833) CELLINK
1
BIO X Autocalibration Kit
Installation and Quick-start Guide
The Autocalibration Kit enables Automatic Bed Leveling (ABL) and automatic X, Y and Z
calibration. These features are available on HeartOS software version 1.7 and above.
The BIO X autocalibration removes the potential for user error when calibrating the tip of the
nozzle relative to the printing surface or to another nozzle. It does this by probing and aligning
all printhead nozzle tips in all axes (X, Y and Z), and includes ABL.
ABL measures the print bed surface and adjusts all movements to follow its tilt and contours to
improve the quality of bioprinted structures. ABL compensates for any height differences
present in a print bed or printing surface. After a brief calibration procedure, the print bed
adjusts the Z height while running the bioprinting protocol, ensuring the construct is level
relative to the printing surface. ABL requires a surface probe to automatically measure the
print bed’s height at various points.

Boston, U.S.
100 Franklin St.，
Boston, MA 02110
Gothenburg, Sweden
Arvid Wallgrens Backe 20,
Gothenburg, 41346
Virginia, U.S.
2000 Kraft Dr., Suite 2125
Blacksburg, VA 24060
Kyoto, Japan
46-29 Yoshida-Shimo Adachi-
cho, Sakyo-ku, Kyoto
www.cellink.com | +1 (833) CELLINK
2
BIO X Autocalibration Kit parts
Part Quantity
Surface probe
1
Surface probe stabilizer
1
Surface probe magnet
1
Z-station base
1
Z-station plate
1
XY printhead calibration lever
1

Boston, U.S.
100 Franklin St.，
Boston, MA 02110
Gothenburg, Sweden
Arvid Wallgrens Backe 20,
Gothenburg, 41346
Virginia, U.S.
2000 Kraft Dr., Suite 2125
Blacksburg, VA 24060
Kyoto, Japan
46-29 Yoshida-Shimo Adachi-
cho, Sakyo-ku, Kyoto
www.cellink.com | +1 (833) CELLINK
3
M4 screw
1
Tools needed:
1) Hex 2.5 screwdriver.
2) T6 screwdriver.
Before installing the BIO X Autocalibration Kit
NOTE: Before installing the BIO X Autocalibration Kit, make sure that your BIO X is running
HeartOS 1.7 or above. If the HeartOS software installed on the BIO X is version 1.6 or older,
your system will not be compatible with the Autocalibration Kit.
To download the most recent version of HeartOS:
1. Visit dna.cellink.com.
2. Create an account if you haven’t already.
3. Download the HeartOS 1.7 update file. Transfer the file to a USB flash drive.
4. Plug the USB flash drive into the BIO X.
5. Navigate to the Utilities menu and select Updates.
6. Select and install the downloaded HeartOS.
NOTE: Before installing the Autocalibration Kit, verify that the autocalibration endstops are
functional. Follow the steps below:
1. Visually inspect the following endstops:
-The surface probe endstop. This is the red button switch under the print box.
-The Z-probe endstop. This is the metal or plastic switch on the top-right side of the
print bed.
-The X-Y endstops. These are in the hole on the top-left side of the print bed.
2. To check for proper activation of the autocalibration endstops, navigate to the Utilities
menu. Select Printer, Hardware then Sensors.
a. Press your finger on the surface probe endstop. Verify that the status light
turns blue to indicate a status change for the surface probe.

Boston, U.S.
100 Franklin St.，
Boston, MA 02110
Gothenburg, Sweden
Arvid Wallgrens Backe 20,
Gothenburg, 41346
Virginia, U.S.
2000 Kraft Dr., Suite 2125
Blacksburg, VA 24060
Kyoto, Japan
46-29 Yoshida-Shimo Adachi-
cho, Sakyo-ku, Kyoto
www.cellink.com | +1 (833) CELLINK
4
b. Press your finger on the Z probe. Verify that the status light turns blue to
indicate a status change for the Z-probe.
c. Use a cartridge with a nozzle attached to reach inside the hole on the top-left
side of the print bed and activate the X and Y endstops. Verify that the status
light turns blue to indicate a status change for the X-probe and Y-probe.
NOTE: If any endstop components are misaligned, absent or not activating, contact the
CELLINK Support team at sup[email protected].

Boston, U.S.
100 Franklin St.，
Boston, MA 02110
Gothenburg, Sweden
Arvid Wallgrens Backe 20,
Gothenburg, 41346
Virginia, U.S.
2000 Kraft Dr., Suite 2125
Blacksburg, VA 24060
Kyoto, Japan
46-29 Yoshida-Shimo Adachi-
cho, Sakyo-ku, Kyoto
www.cellink.com | +1 (833) CELLINK
5
Installation of BIO X Autocalibration Kit
Step 1: Installing the surface probe station
Step 2: Installing the Z-probe station
NOTE: Be careful not to damage the Z probe endstop during the hardware installation.
Excessive force can cause the lever to detach.
1. Turn the BIO X off and remove all
printheads.
2. Unscrew the bottom mechanical endstop
screw of the second printhead, labeled by
the green circle in the diagram.
3. Remove the spacer and keep the M2 screw,
labeled by the red circle in the diagram.
4. Insert the magnet into the surface probe
stabilizer. Polarity does not matter when
inserting the magnet.
5. Align the surface probe stabilizer and
magnet such that the surface probe
endstop is directly on top of the magnet.
6. Being careful not to bend the red switch,
press the stabilizer up and toward the back
of the printer.
7. Insert the M2 screw in the surface probe
stabilizer. Tighten it, making sure to not
overtighten.
1. Align the slot of the Z-station
base with the right M4 hole in the
print bed.
2. Place the supplied M4 screw into
the hole. Do not tighten.
3. Press the Z-station base firmly to
the left to align it with the print
bed.
4. Maintain pressure and tighten the
M4 screw.
1
3

Boston, U.S.
100 Franklin St.，
Boston, MA 02110
Gothenburg, Sweden
Arvid Wallgrens Backe 20,
Gothenburg, 41346
Virginia, U.S.
2000 Kraft Dr., Suite 2125
Blacksburg, VA 24060
Kyoto, Japan
46-29 Yoshida-Shimo Adachi-
cho, Sakyo-ku, Kyoto
www.cellink.com | +1 (833) CELLINK
6
Step 3: Installing the XY printhead calibration lever
5. Insert the Z-probe plate as shown.
It will click into place and pivot freely,
triggering the Z-probe endstop when
pressed down and resetting when
released.
1. Turn the BIO X on and navigate to the
Utilities
menu. Select
Printer
,
Hardware
then
Sensors.
2. Align the left and right edges of the XY
lever to be parallel with the print bed
edges along the Y-axis
3. Gently push the XY lever into the XY
station slot.

Boston, U.S.
100 Franklin St.，
Boston, MA 02110
Gothenburg, Sweden
Arvid Wallgrens Backe 20,
Gothenburg, 41346
Virginia, U.S.
2000 Kraft Dr., Suite 2125
Blacksburg, VA 24060
Kyoto, Japan
46-29 Yoshida-Shimo Adachi-
cho, Sakyo-ku, Kyoto
www.cellink.com | +1 (833) CELLINK
7
Quick-start guide
Inserting and removing the surface probe
1. Insert the surface probe wide-side-up into the surface probe stabilizer. The magnet will
hold it in place.
2. To remove after ABL or autocalibration, pull it away from the magnet and store it
safely.
Machine calibration
NOTE: Perform machine calibration before using any of the automatic calibration features.
Recalibrate after each software update.
HeartOS 1.7+ directory:
Utilities -> Move and calibrate -> Surface probe -> Calibrate
If any X or Y probe sensors are triggered after
installing the XY lever, follow these steps:
1. Turn the round base of the XY lever one quarter
turn clockwise to trigger the X and Y probe
sensors. Continue turning until X and Y probes
are marked as OFF.
2. Move the XY printhead calibration lever to
trigger the X and Y probes and manually
confirm that they are triggered. The XY lever
should automatically return to its center
position.

Boston, U.S.
100 Franklin St.，
Boston, MA 02110
Gothenburg, Sweden
Arvid Wallgrens Backe 20,
Gothenburg, 41346
Virginia, U.S.
2000 Kraft Dr., Suite 2125
Blacksburg, VA 24060
Kyoto, Japan
46-29 Yoshida-Shimo Adachi-
cho, Sakyo-ku, Kyoto
www.cellink.com | +1 (833) CELLINK
8
1. Make sure the surface probe is attached.
Figure 1: Correct placement of surface probe in the surface probe stabilizer.
2. Navigate to Utilities. Select Move and calibrate.
3. Home the axes and move the ABL probe so that it hovers over the XY printhead
calibration lever. Focus on the X and Y alignments; ignore the Z alignment as it does not
need to touch in the Z direction.

Boston, U.S.
100 Franklin St.，
Boston, MA 02110
Gothenburg, Sweden
Arvid Wallgrens Backe 20,
Gothenburg, 41346
Virginia, U.S.
2000 Kraft Dr., Suite 2125
Blacksburg, VA 24060
Kyoto, Japan
46-29 Yoshida-Shimo Adachi-
cho, Sakyo-ku, Kyoto
www.cellink.com | +1 (833) CELLINK
9
a. Cross-reference with correct and incorrect calibration positions pictured
below.
Figure 2. Correct calibration position.
Figure 3. Incorrect calibration position.
Figure 4 Incorrect calibration position.

Boston, U.S.
100 Franklin St.，
Boston, MA 02110
Gothenburg, Sweden
Arvid Wallgrens Backe 20,
Gothenburg, 41346
Virginia, U.S.
2000 Kraft Dr., Suite 2125
Blacksburg, VA 24060
Kyoto, Japan
46-29 Yoshida-Shimo Adachi-
cho, Sakyo-ku, Kyoto
www.cellink.com | +1 (833) CELLINK
10
4. Select Calibrate. The ABL probe will calibrate automatically.
5. Remove the ABL probe.
Figure 5. Remove the probe when autocalibration is not running.

Boston, U.S.
100 Franklin St.，
Boston, MA 02110
Gothenburg, Sweden
Arvid Wallgrens Backe 20,
Gothenburg, 41346
Virginia, U.S.
2000 Kraft Dr., Suite 2125
Blacksburg, VA 24060
Kyoto, Japan
46-29 Yoshida-Shimo Adachi-
cho, Sakyo-ku, Kyoto
www.cellink.com | +1 (833) CELLINK
11
Enabling automatic bed leveling (ABL) for a print
WARNING. Do NOT leave BIO X unsupervised while it runs ABL. Be prepared to unplug BIO X
if ABL fails. Leaving BIO X unsupervised during the ABL process risks damage to your machine.
HeartOS 1.7+:
Bioprint-> Print -> Print -> Calibrate -> Automatic bed-leveling
1. If you would like to run automatic bed- and surface-leveling only, begin the bioprinting
process as usual. Select a model and surface.
2. Enter your printer and layer settings.
3. Navigate to the Print tab. Review your parameters and optionally save the protocol.
4. Select Print.
5. Once in the print page, press Calibrate and select Automatic bed-leveling.

Boston, U.S.
100 Franklin St.，
Boston, MA 02110
Gothenburg, Sweden
Arvid Wallgrens Backe 20,
Gothenburg, 41346
Virginia, U.S.
2000 Kraft Dr., Suite 2125
Blacksburg, VA 24060
Kyoto, Japan
46-29 Yoshida-Shimo Adachi-
cho, Sakyo-ku, Kyoto
www.cellink.com | +1 (833) CELLINK
12
6. A message will appear prompting you to attach the probe. Attach the probe and select
Continue.
7. A progress screen will appear. If necessary, press Abort to cancel the procedure at any
time. After canceling the procedure or completing the ABL process, the system will
return to the calibration method menu.
8. Use automatic axis calibration or remove the probe for manual calibration.
NOTE: Keeping the probe attached while printing may damage the printed construct.

Boston, U.S.
100 Franklin St.，
Boston, MA 02110
Gothenburg, Sweden
Arvid Wallgrens Backe 20,
Gothenburg, 41346
Virginia, U.S.
2000 Kraft Dr., Suite 2125
Blacksburg, VA 24060
Kyoto, Japan
46-29 Yoshida-Shimo Adachi-
cho, Sakyo-ku, Kyoto
www.cellink.com | +1 (833) CELLINK
13
Enabling automatic calibration for a print
WARNING. Do NOT leave BIO X unsupervised while it runs automatic calibration. Be
prepared to unplug BIO X in the event of automatic calibration failure. Leaving BIO X
unsupervised during the automatic calibration process risks damage to your machine.
NOTE: Automatic calibration is only supported for the 3cc Pneumatic, 10cc Pneumatic and
Temperature-controlled Printheads.
HeartOS 1.7+:
Bioprint-> Print -> Print -> Calibrate -> Automatic Calibration
1. If you would like to run automatic bed-leveling and X, Y and Z axis calibration, start the
bioprinting process as usual. Select a model and a surface and enter your printer and
layer settings.
2. Go to the Print tab. Review your parameters and optionally save the protocol.
3. Select Print.
4. Press Calibrate in the print page. Select Automatic calibration.

Boston, U.S.
100 Franklin St.，
Boston, MA 02110
Gothenburg, Sweden
Arvid Wallgrens Backe 20,
Gothenburg, 41346
Virginia, U.S.
2000 Kraft Dr., Suite 2125
Blacksburg, VA 24060
Kyoto, Japan
46-29 Yoshida-Shimo Adachi-
cho, Sakyo-ku, Kyoto
www.cellink.com | +1 (833) CELLINK
14
5. A message will appear prompting you to attach the probe. Attach the probe and select
Continue.
6. Ensure the XY probe is positioned in a straight vertical position. Clear the bed of any
tall objects.
7. Verify all hardware components are ready.
8. Select Start to begin the calibration process.

Boston, U.S.
100 Franklin St.，
Boston, MA 02110
Gothenburg, Sweden
Arvid Wallgrens Backe 20,
Gothenburg, 41346
Virginia, U.S.
2000 Kraft Dr., Suite 2125
Blacksburg, VA 24060
Kyoto, Japan
46-29 Yoshida-Shimo Adachi-
cho, Sakyo-ku, Kyoto
www.cellink.com | +1 (833) CELLINK
15
9. After a homing cycle, automatic calibration will begin and a progress screen will appear.
If necessary, press Abort to cancel the procedure at any time.
10. After the automatic calibration process is complete, a message will appear prompting
you to remove the probe. Remove the probe and select Close to return to the print
menu.
11. Press Start to proceed to bioprinting.

Boston, U.S.
100 Franklin St.，
Boston, MA 02110
Gothenburg, Sweden
Arvid Wallgrens Backe 20,
Gothenburg, 41346
Virginia, U.S.
2000 Kraft Dr., Suite 2125
Blacksburg, VA 24060
Kyoto, Japan
46-29 Yoshida-Shimo Adachi-
cho, Sakyo-ku, Kyoto
www.cellink.com | +1 (833) CELLINK
16
Removing hardware for packing
When packing BIO X for transport, remove the following parts:
•XY printhead calibration lever.
•Z station base.
•Surface probe.
Tools are not required to remove these parts. Ensure they are stored safely during transport.