Cellink Inkredible+ User manual

User Manual
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“You cannot create new science unless you
realize where the old science leaves off and
new science begins, and science- fiction
forces us to confront this.”
Michio Kaku

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Table of Contents
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1
INTRODUCTION OF
YOUR INKREDIBLE
3D BIOPRINTER

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INTRODUCTION OF THE INKREDIBLE
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Hello and welcome to the exciting world of 3D Bioprinting. The
technology you have in front of you has the potential of creating
real, living human life and it is only your imagination that limits
what you can do. This manual will serve as your guide for setting
up your INKREDIBLE bioprinter, maintain it, and also notify you
about safety precautions. Let`s begin this exciting journey!
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WARNING: The INKREDIBLE includes moving parts that can cause injury. Never
reach inside the INKREDIBLE while it is in operation. Always allow the
INKREDIBLE to come to a complete stop prior to reaching inside.
WARNING: Do not leave the INKREDIBLE unattended during operation.
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SPECIFICATIONS
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PRINTING
Print Technology: Extrusion based bioprinting
Build Volume: 130 x 80 x 50 mm
High 100 microns [0.0039 in]
Standard 200 microns [0.0078 in] Low 300 microns [0.0118 in]
Positioning Precision: XY: 11 microns [0.0004 in]; Z: 2.5 microns [0.0001 in]
Needle Diameter: User dependent
Needle Height: User dependent
SOFTWARE
Software Bundle: Repetier Host
File Types: .stl, .obj, .thing
Supports: Windows (XP 32 bit/7+)
Ubuntu Linux (12.04+)
Mac OS X (10.6 64 bit/10.7+)
PHYSICAL DIMENSIONS
19.1 x 16.5 x 14.7 in [330 x 370 x 300 mm]
Shipping Box: 23 x 21.5 x 17 in [59 x 55 x 43 cm]
Weight: 25.4 lbs [11.5 kg]
Shipping Weight: 44.55 lbs [20.2 kg]
ELECTRICAL
AC Input: 100 – 240 V, ~2 amps, 50 – 60 Hz
Power Requirements: 24 V DC @ 6.25 amps
Connectivity: USB, SD card [FAT 16, max. 2GB]
MECHANICAL
Body: Powder-coated high grade steel
Build Platform: Acrylic
XYZ Bearings: Wear-resistant, oil-infused steel
Stepper Motors: 1.8° step angle with 1/16 micro-stepping

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HOW DOES IT WORK?
The INKREDIBLE bioprinter is an extrusion based 3D Bioprinter with dual extruder
heads for bioprinting of human tissue models and organs. It is a cost-effective unit to
enter the 3D bioprinting field and for innovators and beginners to start bioprinting living
tissues at ease. It can be used as a standalone unit thanks to its LCD display or it can
be monitored through a computer with the use of the accompanied software. Your
three-dimensional design files are translated into coordinates and instructions for the
INKREDIBLE and allows the bioprinter to move according to a defined plan. The
printing process works through the extrusion of a bioink or hydrogel material combined
with the extrusion of human cells layer by layer until a 3D structure is built from bottom
to top. Once the structure has been printed, it can be cross-linked using the Blue LED
UV crosslinker or ionic solutions, depending on your bioink material.

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2
UNPACKING YOUR
INKREDIBLE 3D
BIOPRINTER
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UNPACKING YOUR INKREDIBLE 3D BIOPRINTER
When unpacking your INKREDIBLE 3D Bioprinter, be careful not to grab onto any
movable parts or lose components. We recommend you grab it by the top window frame
and pull it out from the box.
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NOTE: Do not force or tear anything during the unpacking or setup process as
this may irreversibly damage your INKREDIBLE 3D Bioprinter
CAUTION: When opening the box, do not force or tear anything while
unpacking as this may damage the INKREDIBLE 3D Bioprinter.

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CONTENT OF THE BOX
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[1x] INKREDIBLE 3D Bioprinter
[1x] 24V Power supply
[1x] Power cord
[1x] SD card pre-loaded with sample objects to print
[1x] USB Cable
[1x] Portable oil-free air compressor with tubing
[1x] STARTINK-Kit (1 prefilled Syringe with 3mL of CELLINK + 1 CELLMIXER + 10mL
of Crosslinking solution)
[3x] Empty cartridges
[1x] 24 well plate
[1x] Petri-dish
Note: Unpacking should be done outside the laboratory environment.

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INKREDIBLE DIAGRAM
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NOTE: The INKREDIBLE Bioprinter is coming in one single piece and SHOULD NOT
be taken apart.
The dual printheads are made to fit Nordson EFD 3mL cartridges.
The pressure pump has a maximum pressure to be set of 7 bars and should be set at
2 bars minimum to have the extruder heads functioning properly.
[1] On/Off Button
[2a] Printhead 1 (Ph 1)
[2b] Printhead 2 (Ph 2)
[3a] Digital Pressure Monitor for Printhead 1
[3b] Digital Pressure Monitor for Printhead 2
[4] LCD Screen
[5] Control Knob
[6] UV Crosslink LED
[7] Printbed
[8a] Pressure Regulator for Printhead 1
[8b] Pressure Regulator for Printhead 2
[9] Connections
[10] X-Axis Stepper Motor

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SAFETY INSTRUCTIONS
MALOPERATION OF THE INKREDIBLE 3D BIOPRINTER
INSTRUMENT CAN LEAD TO SEVERE AND IRREVERSIBLE
INJURIES IF THE SECURITY INSTRUCTIONS ARE NOT
FOLLOWED!
NOTE: In order to respond to the INKREDIBLE 3D Bioprinter and its relevant accessories
security requirements, the instrument has to be operated by trained personnel, which is aware
of the risks and dangers that can be induced to themselves, as well as for the staff being in
close relation with the INKREDIBLE 3D Bioprinter.
The staff being involved in the installation, or the maintenance of the system or a part of the
system, has to be qualified through an appropriate training.
GENERAL SAFETY INFORMATION
Use the instrument only for its intended purpose as described in the documentation. Do not
modify the instrument, sub-components or accessories.
Never reach into the instrument until all components of the instrument have
come to a complete stop. Moving parts can cause serious injuries.
Before cleaning, inspecting, adjusting, repairing or disassembling the
instrument or the dispensing units, stop the instrument and turn off the main
switch.
The INKREDIBLE Bioprinter is fitted with a 365nm (405nm option)
UV LED. DO NOT LOOK DIRECTLY AT THE LIGHT. ALWAYS
WEAR PROTECTIVE EQUIPMENT WHEN OPERATING THE
SYSTEM. ALWAYS WEAR THE INCLUDED SAFETY GLASSES.
Improper mounted pressurized air spouts, liquid spouts or cartridges can be
dangerous. Make sure all connections and connectors are mounted
according to instructions and all components are specified for use with the
intended pressure.

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NOTE: Make sure the supplied air pressure does not exceed the admissible range of 4 bars
(400 kPa).
PERSONNEL PROTECTIVE EQUIPMENT
Always wear protective glasses and secure the environments of the instrument to other
personnel while operating the INKREDIBLE. All laser safety equipment, including laser safety
glasses, must be adapted to the installed UV curing kit.
Always wear protective glasses and gloves while handling dangerous materials that can be
absorbed by skin or which are toxic, strike-attractive, corrosive, causing allergies, carcinogen,
reproduction-endangering or mutagenic to human being.
Always read the material safety data sheets, packing labels, manufacturer or dealer’s catalogue
first to make sure to use adequate safety equipment.
LIQUIDS, DISPENSING MEDIUMS
Before starting to work with hazardous dispensing materials the
operator has to inform himself about the characteristics of the
material and its compatibility with the dispensing unit. The
information can be drawn from the material safety data sheets,
packaging labels, manufacturer or dealer catalogues.

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3
SETUP &
INSTALLATION
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OPERATING THE INKREDIBLE
Only trained staff is allowed to operate the instrument being aware of the risks and potential
injuries (non reversible damages can be caused).
NOTE: OPENING OF THE INSTRUMENTS ELECTRICAL PANEL WILL AUTOMATICALLY
REVOKE THE WARRANTY.
The manufacturer certifies that the INKREDIBLE is conforming to requirements.
The manufacturer gives an 8 months warranty on the moving parts in the INKREDIBLE
bioprinter. The guarantee excludes deliberate damages.
SETUP AND INSTALLATION
1. Unpacking the 3D Bioprinter
Once the INKREDIBLE 3D Bioprinter has been carefully taken out of the box, all protective films
plastic parts preventing the axis to move during transportation can be removed. Once all
protective materials and films have been removed, the INKREDIBLE can be placed inside a
biological hood or laboratory bench. The INKREDIBLE should never be placed on
wet/damp/moist/humid surfaces. Make sure to remove all BLACK zip ties holding the X-
carriage and Printbed in place during transportation.
2. Remove the Accessory Box and the Air Supply Box
3. Place the INKREDIBLE bioprinter inside the biological hood
4. Calibrating the printbed for the first time (every time you move the INKREDIBLE you
should follow this calibration procedure):
A standard leveler should be used for calibrating the printbed for the first time.
Place the leveler at the back of the printbed and adjust the back screws to level.
Place the leveler on the middle of the printbed and perpendicular to the back screws. Adjust the
front screw to level.
5. The INKREDIBLE 3D Bioprinter can be placed inside an isolator
6. The power cord can be plugged in the back [9] of the of the INKREDIBLE 3D bioprinter
and then in the outlet inside of the isolator or outside the isolator. Note: Ensure to always
plug in the power cord into a separate individual outlet for bioprinter.

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7. The INKREDIBLE 3D bioprinter can be switched on with the on/off button [1].
8. Connect the air tube (provided) to the air compressor and then to the back of the
Bioprinter. Turn on the air compressor and regulate the air pressure to 400 KPa. Note:
Air compressor should be connected to a separate socket (not in the same fuse) to avoid
any electromagnetic interference with the INKREDIBLE Bioprinter. Air pressure should
be set at minimum 200 KPa to have the printhead’s actuators functioning properly.
9. Connect the cartridges with needles to air tubes for Print Heads [2a] [2b] by twisting the
cartridge connectors.
10. Attach the cartridges connected to the air tubes inside the Printheads [2a] [2b].
11. Start off by homing and calibrating the INKREDIBLE bioprinter. The bioprinter can be
calibrated and tuned using the “Home Axes” function in the menu, using the following
sequence (Note: Ensure that the cartridges with needles are placed inside the
printheads prior to initiating this sequence):
To Home Axes the bioprinter:
-Remove all objects from the build plate [7] ( such as well plates, petri dishes etc. )
-Lower the build plate [7] by simply pushing down at the back of the printbed with your hand
-Press the Control Knob [5] once to enter the “Menu”
-Turn the Control Knob Wheel [5] to select “Prepare Bioprint” and press the Control Knob to
enter
-Turn Control Knob wheel and press “Home Axes”
The INKREDIBLE Bioprinter will now home all axes. The Ph 1 needle should in the middle of
the printbed once the sequence is completed.
Manual adjustment and Calibration of the Z Axis to print in petridish and well plates (should be
performed right after “Home Axes” sequence):
-Place a petridish onto the Build Plate [7] (there are cuts in the center of the build plate where a
petri dish will fit tightly)
-Press the Control Knob Wheel [5] once to enter the “Menu”
-Turn the Control Knob Wheel [5] to select “Prepare Bioprint” and press the Control Knob to
enter
-Turn the Control Knob Wheel to select “Move Z” and press the Control Knob You have now
entered Manual Z Calibration Mode-On the display you should see:Z: +040.0 (position of Z
axis: 40 mm)

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-Turn the Control Knob Wheel [5] until the Ph 1 needle is right above the petri dish or well plate
well.
A little space of about 0.1mm should be visible between the print bed and the needle tip. You
can use a piece of paper to slide between the needle and the petri dish to measure the
distance. The paper should slide freely but there should not be more space than that.
-Once the correct distance has been selected, Press the Control Knob Wheel [5] once to exit
the Manual Calibration Z mode
-Scroll the Control Knob Wheel and select “Calibrate Z”
Your INKREDIBLE 3D Bioprinter is now calibrated and ready to print!
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STARTING THE INKREDIBLE IN STAND-ALONE MODE
-Turn on the INKREDIBLE +, followed by the Air Compressor.
-Lock the air tube onto the sample cartridge.
-Clip the cartridges onto Printhead 1 (Ph 1) and Printhead (Ph 2)
-Follow the procedure to “Home Axes” and “Calibrate Z” in the INKREDIBLE
-Clip the 24 well plate or petri dish onto the printbed
- Adjust the pressure using the two separate pressure regulators located on the right side of the
INKREDIBLE 3D Bioprinter. Note: the regulator closest to the user is for Printhead 1 and the
one further away is for Printhead 2.
-In the LCD menu, select “Bioprint from SD” and then select the file
-The bioprinting process is now running! The pressure can be adjusted during bioprinting in
order to have a continuous flow, simply turn on the pressure regulators.
-At the end of the bioprinting, the printbed will lower.
-Take out the well plate or petri dish.
-To Crosslink with the UV System, enter the “Utilities Menu” on the control panel by pressing the
Control Knob Wheel [5].
-Select “Utilities” and press the Control Knob [5]
-Scroll until you select “Turn On LED” and press the Control Knob button. To turn off the UV
LED curing, press the “Turn Off LED” button.
You can try bioprinting any of your bioinks or hydrogels. Simply, load it in the cartridges and
configure your bioprinting parameters (Printing pressure and printing speed). Configurations,
pressure adjustments, and any development of printable biomaterial is upon the customer`s
responsibility. CELLINK is not responsible for any clogging or damages caused by any user
dependent biomaterials bioprinted with the INKREDIBLE.
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BIOPRINTING WITH CELLINK SOFTWARE (REPETIER
HOST)
INTRODUCTION TO G-CODE PROGRAMMING
The special M codes for INKREDIBLE 3D Bioprinter
M712 to M732 are used to set the offset between the printheads
printhead 1 and 2 are the syringes and printhead 3 is the led.
Note: That this will not place the chosen printhead in the active position.
M712 Set offset from 1 to 2
M713 Set offset from 1 to 3
M721 Set offset from 2 to 1
M723 Set offset from 2 to 3
M731 Set offset from 3 to 1
M732 Set offset from 3 to 2
M750 homes the printheads. This needs to be done before any automatic switching of the
printheads can occur. After this command Printhead 1 will be in the active position
M751 Places Printhead 1 in the active position. This is faster than M750.
M752 Places Printhead 2 in the active position
M753 retracts both Printhead 1 and 2 making way for the led to be used.
M760 Opens the valve for printhead 1
M761 Closes the valve for printhead 1
M762 Opens the valve for printhead 2
M763 Closes the valve for printhead 2
M764 Turns on the UV LED
M765 Turns off the UV LED
Workflow
Import the models into the slicing software and slice.
Open the G-code file and make sure to remove any homing command in the start of the code.
INKREDIBLE needs to be homed manually from the menu to account for different print heights.
NOTE: FOLLOW THE CALIBRATE PRINTBED SEQUENCE ABOVE.