Allevi 1

User Guide

Allevi 2
User Guide – 1.5
BUILD WITH LIFE

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Table of Contents

WELCOME 4

SAFETY AND COMPLIANCE 5

SPECIFICATIONS 5

SETTING UP YOUR ALLEVI 7

WHAT’S IN THE BOX 7

DIAGRAMS 8
GETTING STARTED – INSTALLING SOFTWARE 9
GETTING STARTED – SETTING UP PRINTER 11

YOUR FIRST BIOPRINTS 14

PRINTING SINGLE MATERIAL STRUCTURES 14

PRINTING TWO MATERIAL STRUCTURES 25
PRINTING THERMAL PLASTICS 31

MAINTENANCE & TROUBLESHOOTING 33

RESETTING PASSWORDS 33
TROUBLESHOOTING AND CONTACT US 34

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This User Guide is designed to help start your journey
into the deep unknowns of 3D bioprinting.
In the first section of this guide you will learn about the
basic components of your Allevi, how to set it up,
download the correct software and load bioinks.
The second section of this guide will provide you with
instructions on using the Allevi software to print a single
material structure
The third section will teach you how to print a dual
material structure.
Finally the fourth section will teach you how to print
thermal plastics using the heating components.
Following this guide will help ensure that you are getting
the most out of your machine. We can’t wait to see what
you will create.
SPECIFICATIONS

Printing Electrical
Print Technology: Fused Deposition Power Requirements: 6Amp AC Desktop
Manufacturing Adaptor
Build Volume: 9 x 9 x 9 cm (729 Connectivity: USB
cm3 ), 3.5 x 3.5 x 3.5 inch (42.87 in3 )
Software
Layer Resolution: 100 micron
Bundle: Modified Repetier Host
Nozzle Diameter: 100 micron
Supported File Types: 3D STL
Print File Type: GCODE
Operating System: Windows, Mac OS X,
Linux
Mechanical
Construction: Aluminum Frame
Extruders
Build Structure: Tissue Culture Plates
Max Temp: 120 Degrees Celcius
Stepper Motors: 0.9 Degrees Step
LED Range: 400-410 nm
Angle
XY Position Precision: 5.5 microns
Z Position Precision: 5 microns

WARNING: The Allevi 1 includes moving parts that can cause injury. Never reach inside
the printer while it is in operation.
WARNING: Do not leave the printer unattended while it is in operation.
WARNING: Always wear safety goggles when using the printer.
WARNING: Extruders heat to high temperatures. Always allow time for extruders to cool
before handling.

CAUTION: Always power off the printer before reaching inside.

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Let’s Get Started!
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WHAT’S IN THE BOX

Allevi 2 Printer Safety Goggles USB Cable

Power Adapter Air Compressor Compressor Hose

Sample Bio Kit

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ALLEVI 2 DIAGRAM

A. Air Compressor Lines

B. Dual Extruder
C. Build Plate
D. Power On/Off

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GETTING STARTED

Installing the Software

1. First install Repetier Host:
https://drive.google.com/file/d/0BxscXTKIWFFEUGs2bGhxak04anc/view?usp=sh
aring

This will install Python 2.7 and an FTDI Driver. Close window if Repetier-Host
Server opens in your browser and continue.

2. Open Repetier-Host “Printer Settings” and assign the following values.

3. Downloand the Slic3r Config Bundle File:

https://drive.google.com/file/d/1tYVGr5i8uVWdJjkZQg7Zzl5ugwVhBdcL/view?usp=sh
aring

4. Open Slic3r and click “Configuration

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5. Go to File > click “Load Config Bundle…”

6. Install Bioprint Software from this site

Windows

1. Install Python 2.7.

2. Go to and install this before proceeding http://aka.ms/vcpython27
3. Install FTDI Drivers
4. Download Bioprint and extract to desired location on your computer.
5. Navigate to bioprint-master/Windows folder on your file system.
6. Right-click Install.bat file and "Run As Administrator".
7. Double-click Bioprint.bat and leave the Command Prompt Window open. If you
receive an error, press any key to close the window and try
running Bioprint_firewall.bat.
8. Navigate to bioprint/ in your browser if you ran Bioprint.bat or bioprint:9999/ if you
ran Bioprint_firewall.bat
9. Setup a username and password to use with Bioprint and then proceed to register
your printer. Enter in your email address and put the name of your institution in as
the Serial Number. Finally select “Keep Access Control Enabled”
10. Login using the login menu in the top right corner of the screen
11. Begin printing with life!

Mac OSX

1. Install Python 2.7.

2. Download Bioprint and extract to desired location on your computer.
3. Navigate to bioprint-master/MacOSX folder on your file system.
4. Double-click Install.command and enter your Administrator password when
prompted. Press Return to reboot when prompted.
5. After rebooting, download and install FTDI Drivers
6. Double-click Bioprint.command and leave the Terminal Window open to run
Bioprint.
7. Navigate to bioprint/ in your browser.
8. Setup a username and password to use with Bioprint and then proceed to register
your printer. Enter in your email address and put the name of your institution in as
the Serial Number. Finally select “Keep Access Control Enabled”
9. Login using the login menu in the top right corner of the screen
10. Begin printing with life!

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NOTE: For the Mac version you will have to type in your password and reboot your
computer
NOTE: It sometimes takes time to install so let it run until finished.

7. Leave the Command Prompt Window open. Double click to open the Bioprint file. If you
receive an error, press any key to close the window and try running Bioprint_firewall.bat

8. Open your browser (preferably Chrome) and type “bioprint/” into the navigation bar of
your browser. Alternatively type “bioprint:9999/” if your ran Bioprint_firewall.bat

9. Setup a username and password to use with your software. Your user name and
password are computer specific so the one you create will always have to be used for
that computer. You can use the same username, password, and serial number multiple
times on different computers

TroubleShoot: Sometimes when you type in bioprint/ or bioprint:9999/ on the web

browser nothing appears. When this happens open command prompt in the start menu.
In command prompt type “ipconfig”, Find the IPv4 number. Navigate back to your
browser and type this number followed by 9999 into your navigation bar to access
Bioprint.

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Setting up your printer
1. Carefully remove your printer from the packaging by grasping the printer frame
firmly with both hands.

2. Plug the USB cable into the port at the back of your printer and connect the other
end to a USB port on your computer.

3. Use the power cable to connect the printer to a power supply via the port located
on the right side of the printer.

4. Power on the printer by pressing the “On/Off” button located front of the printer

5. Attach the air compressor to the printer by connecting the compressor hose to
nozzle on the left side of the printer. This can be a tricky maneuver until you get
the hang of it. For more in depth directions on connecting the air compressor,
click here to watch a quick video.

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BIOPRINTING WITH ONE MATERIAL

Slicing One STL

You can design STL files on either Solidworks or Autodesk Fusion 360. Alternatively, you can
find a host of STL files in the getting started folder. To begin we will use this predesigned STL
file.

Pediatric-Bronchi.STL:

1. Upload the Pediatric-Bronchi.STL file into Repetier-Host.

a. Move the object on the bedplate using the arrows highlighted on the left.
b. Save, copy, or scale object with the items on the right.
c. Assign the STL to an extruder by using with the extruder drop down menu.

2. Enter your printer settings.

a. Select the “Slicer” tab to assign preconfigured Slic3r settings to slice the file.
b. Speed and layer height are defined.

3. Select the configuration tab to adjust settings important for bioprinting.

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a. Layer Height – Determines how thick or thin your layers are

b. Speed – Determines how fast the extruder will move during the print. Remember to
change all the speeds in the column to the desired number
Recommendation: 2-4 mm/s is good for a small accurate design(example: lattice). 8-16
mm/s is good for a large design(example: ear)

c. Nozzle Diameter – Enter the number that accurately depicts your nozzle tip inner
diameter

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 Note: Your layer height has to be numerically less that your nozzle diameter to correctly
slice
Note: Once the changes have been made remember to save and label your settings.

4. Once your settings have been saved, you are now ready to slice the file.

5. You can view the printer movement by viewing the g-code created by your slicer file. Note: If
no g-code is generated, check that your nozzle diameter is thin enough.

6. Save your g-code file.

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 Printing One STL

1. Navigate to the Bioprint Window and log in.

2. Navigate to the Bioprint window and Upload the G-code file. Note: The Bioprint.bat file must
be loaded and the command window must be kept open in the background.

Note: If you click the wrench you can sort out your gcode files by name or date

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3. Connect your printer to your computer

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4. Home Z

5. Home E

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6. Home X/Y

7. Click “Select Well Plate” Use “Petri Dish”.

Loading Material Into Syringe

For this Print we will use Pluronic F127, please place a bit of this into the fridge to allow it to
become liquid. Note: Different materials will be loaded differently depending on the type of
material they are.

Watch this video to learn how to prepare Pluronic to print with your Allevi 2. We suggest filling
your syringe with about 3 ml of material for your first print:
https://www.youtube.com/watch?v=AZYw5u53PiM

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Calibrating Extruder
1. First place cap on syringe

2. Load syringe into canister and tighten knob.

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3. Now set and test pressure for extrusion. Set pressure by manually increasing the
settings using the +/- buttons (image A). Test the pressure by pressing extrude (image
C). Adjust pressure between approximately 50-80 PSI for pluronic until material
extrudes continuously (image D).
A B

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4. Switch step size from 10mm to 1mm. Then use Software to move the bed plate (Z)
closer to the Extruder(Extruder 1) into the plate to the point where the dish is almost
touching the needle. Finer 0.1mm movement can be used when approaching the dish.

5. Once the Z axis is in place, press “extrude” for extruder 1 to turn the pressure on and
deposit a small amount of material, then quickly press “extrude” again to turn the
extruder off.

NOTE: Sometimes the syringe moves slightly when you hit extrude. If it is too low it will not
create a good dot and no material will come out. If it is too high then a dangling line will
be created.

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6. Once you have the correct orientation, click “Update” (image A) under extruder 1, then
“Calibrate” (image B).
A B

7. Now that your tool is calibrated, you are ready to print!! When ready, click Load and
Print

8. Sit back and marvel at your awesome first bioprint!

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BIOPRINTING WITH TWO MATERIALS

Slicing Two STL Designs

FOR MAC: Open Slic3r and follow this video.
FOR WINDOWS:
1. Import two STL designs and click “Center Object” to align them both
2. Find the two downloads here:
3. Ring Support Ring Infill

4. Assign each STL to an extruder using the Extruder drop down menu.

5. Once desired Slic3r settings are configured. Click “Slice with Slic3r”.

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 6. Save the G-Code created.

Printing Two STL Designs

1. Load your two separate syringes with Pluronic.

2. Connect your Allevi 2 to your computer.

3. Home Z. Home E. Home X/Y.

4. Select “Petri Dish” from the “Select Plate” Type menu. The click “Select well plate”.

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 1. Calibrate pressures of each extruder. Set pressure by manually increasing the
settings using the +/- buttons (image A). Test the pressure by pressing extrude
(image B). Adjust pressure between approximately 50-80 PSI for pluronic until
material extrudes continuously (image C).
A B

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5. Switch step size from 10mm to 1mm. Then use Software to move the bed plate (Z)
closer to the Extruder(Extruder 1) into the plate to the point where the dish is
almost touching the needle. Finer 0.1mm movement can be used when
approaching the dish.

9. Once the Z axis is in place, press “extrude” for extruder 1 to turn the pressure on and
deposit a small amount of material, then quickly press “extrude” again to turn the
extruder off.

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10. Once you have the correct orientation, click “Update” (image A) under extruder 1, then
“Calibrate” (image B).
A B

6. Now use the extruder controls to bring the bed plate up to the second extruder so
the needle is almost touching the dish. Next, use X/Y controls as well to line up the
right extruder directly on top of the dot you created with the left extruder.

7. Once you have the correct orientation, click “Update” under extruder 2, then
“Calibrate.”

8. Load your dual print G-Code file.

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9. Click “Load and Print” G-Code

10. Pat yourself on the back! You’re on your way to becoming a 3D bioprinting master!

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BIOPRINTING WITH PCL
1. Download and import this print into Repeteir-Host:
https://drive.google.com/file/d/1m0rBUzfAd_lkgmdbLfl8_vJTjGmkMiMM/view?usp=sharing

Should you want the specific slicing profile we used you can find those here too:
https://drive.google.com/file/d/18WLSq_Gm5mOq1MiA3nic3b_jyyUVvKV4/view?usp=sharing

2. This time you will need your metal syringe and a metal tip for use with thermal plastics.

3. Pour the PCL beads directly into the metal syringe.

4. Load the syringe into extruder 1.

5. Set the temperature to the desired target temperature. In this case, we will set the
temperature to 100 degrees Celsius and the pressure to 100 PSI. Allow the extruder time to
heat up.

6. Allow the extruder to heat up. Set the pressure to 100 PSI.

7. Home Z,E,X/Y. Select “Petri Dish” and create a dot onto the dish.

8. Press print and then call your colleagues into the room so you can show off your new skills!

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 TO LEARN MORE VISIT
BUILDWITHLIFE.COM
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PASSWORD RESET

Resetting BioPrint Password On Windows

1. Navigate to “C:\Users\<username>\”
2. Unhide hidden items to view the “AppData” folder
a. As depicted below, navigate to “View” and check the box next to “Hidden Items”

3. Navigate into “…\AppData\Roaming\”

4. Delete the BioPrint Folder

Connection Issues On Mac

If you are having trouble with reconnecting your printer on a MAC.

You need to make sure to

1. Disconnect the printer in the software
2. Unplug the printer from your computer
3. Turn the printer off

What is most important here is to make sure that you follow these instructions in this order
because there is actually a known bug in the way that MacOSX handles when serial ports are
disconnected incorrectly and this can cause the serial port to not be detected again when
plugged back in, which is what causes this error.

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Resetting BioPrint Password On MAC
1. Open Finder and select "Go to Folder" from the "Go" menu in the menubar

2. Enter `/Users/[your computer username]/Library/Application Support` as the folder to go

to. If your computer username is Allevi you will enter `/Users/Allevi/Library/Application
Support` here

3. Delete the BioPrint folder

4. Restart BioPrint

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 TROUBLESHOOTING

For problems, questions, or concerns

please visit www.Allevi.io or email us
at support@Allevi.io

Allevi Inc | 3401 Grays Ferry Ave | Building 176 | Philadelphia PA | 19146

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