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Allevi 1 User Guide

Allevi 2 User Manual – 1.6.4

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B U I L D W I T H L I F E

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Table of Contents

WELCOME 4

SPECIFICATIONS 5

SAFETY AND COMPLIANCE 5

GETTING STARTED 6

WHAT’S IN THE BOX 7

ALLEVI 2 DIAGRAM 8

SETTING UP YOUR PRINTER 9

SETTING UP YOUR SOFTWARE 10

YOUR FIRST BIOPRINTS 13

PRINTING SINGLE MATERIAL STRUCTURES 14

PRINTING DUAL MATERIAL STRUCTURES 24

MAINTENANCE & TROUBLESHOOTING 30

RESETTING PASSWORDS 30

TROUBLESHOOTING AND CONTACT US 32

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This User Manual will help get you started using your Allevi

bioprinter.

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 bioprint for

your desired application.

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!

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SPECIFICATIONS

WARNING: The Allevi 2 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.

Mechanical

Construction: Aluminum Frame

Build Structure: Tissue Culture Plates

XYZ Position Precision: 5 microns

Dimensions: 12 x 12 x 12 in

30.5 x 30.5 x 30.5 cm

Electrical

Power/Voltage Requirements: 6Amp AC

Desktop Adaptor

Connectivity: USB

Software

Bundle: Modified Repetier Host

Supported File Types: gcode

Operating System: Windows, Mac OSX, Linux

Printing

Print Technology: Fused Deposition

Manufacturing

Layer Resolution: 100 microns

Print File Type: Gcode

Build Volume: 5.1 x 3.5 x 2.4 in (42.8 in3)

13 x 9 x 6 cm (702 cm3)

Extruders

Max Temp (E1): 160°C

Max Temp (E2): 70°C

Min Temp: Room temperature

Pressure Range: 0-120PSI (0-820kPa)

LED Range: 405 nm

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Let’s Get Started!

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WHAT’S IN THE BOX

Allevi 2 Printer USB Cable

Power Adapter Starter Bioink Kit

Air Compressor* Compressor Hose

*Air compressor arrives separately

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ALLEVI 2 DIAGRAM

A. Air Compressor Lines

B. Dual Extruder (Left: Extruder 1, Right: Extruder 2)

C. Build Plate

D. Power On/Off

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GETTING STARTED

Setting up your printer

1. Carefully remove your printer from the packaging by grasping the printer frame firmly with

both hands.

2. Use the power cable to connect the printer to a power supply via the port located on the

right side of the printer (labelled Step 2).

3. Power on the printer by pressing the power button located front of the printer.

4. Attach the air compressor to the printer by connecting the compressor hose to nozzle on the

left side of the printer (labelled Step 4). This can be a tricky maneuver, so for more in-depth

directions on connecting the air compressor, watch this quick video. The air compressor

must remain turned on for the entirety of the print. We recommend keeping the air

compressor set to a pressure significantly higher than your print pressure. This is so that as

the air compressor loses some pressure and refills, your print is not affected. The air

compressor’s pressure can be adjusted by turning the black knob. It has a max pressure of

120PSI.

Step 4 Step 2

https://www.youtube.com/watch?v=yQSNN1tncoo

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Setting up your software

The following steps will take you through the software installation process. This process is also

available in this video. If you are a Linux user, please email support@allevi3d.com for

instructions.

1. Download and install FTDI Drivers.

a. Windows: FTDI Drivers (Right-click to “Run As Administrator”)

b. MacOSX: FTDI Drivers.

2. Download the installer for your operating system from the Allevi Software page.

3. Double-click desktop application to run Bioprint.

4. The software will open in your browser. To create an account:

a. Choose a username and password to use with Bioprint

b. Enter your email.

c. For the ‘Serial Number’ field, put the name of your institution or lab.

d. Finally select “Keep Access Control Enabled.”

5. Login using the login menu in the top right corner of the screen.

6. To close the software, quit from the taskbar.

NOTE: Your username and password are computer specific so the one you create will always have to be used for that computer.

https://www.youtube.com/watch?v=iWV0Lfva3womailto:support@allevi3d.comhttp://www.ftdichip.com/Drivers/CDM/CDM%20v2.12.12%20WHQL%20Certified.exehttp://www.ftdichip.com/Drivers/VCP/MacOSX/FTDIUSBSerialDriver_v2_3.dmghttps://allevi3d.com/software

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7. Install Repetier Host.

Close window if Repetier-Host Server opens in your browser and continue.

8. Open Repetier-Host. Go to “Config” > “Printer Settings” and assign the following values.

9. Download the Slic3r Config Bundle file.

10. In Repetier Host, go to the Slicer tab and click “Configuration”

https://www.repetier.com/download-now/https://biobot2.squarespace.com/s/Allevi-Slic3r-Bundle-195.ini

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11. Go to “File” > “Load Config Bundle…” and select the Slic3r Config Bundle file you just

downloaded.

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Let’s Start Printing!

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BIOPRINTING WITH ONE MATERIAL Slicing One STL

You can design STL files on any CAD software, such as SolidWorks or Autodesk Fusion 360.

Alternatively, you can find a host of STL files in the “Getting Started” folder. To begin we will use a

predesigned STL file.

1. Load 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 print settings.

a. Select the “Slicer” tab to assign preconfigured Slic3r settings to slice the file.

b. Speed and layer height are defined.

https://drive.google.com/file/d/1EBHiCZgaenBO-lUV3sVd4mjW74BhFqJH/view?usp=sharing

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3. Select the configuration tab to adjust settings important for bioprinting. Print, filament, and printer

settings are accessible under ‘Settings.’

a. Layer Height – Determines how thick or thin your layers are. This is a property typically

determined by the material type.

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: 2-6 mm/s is good for a small accurate design (example: lattice). 8-16 mm/s is

good for a large design (example: ear). Speed is a print parameter that is very dependent on

material and pressure.

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

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.

a. Save your g-code file.

NOTE: Your layer height must be less than your nozzle diameter to correctly slice NOTE: Once the changes have been made, remember to save and label your settings.

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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: You can sort your gcode files by name, date, or size by clicking the wrench.

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3. Connect your printer to your computer.

4. First, home the Z-axis.

5. Second, home the E-axis.

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6. Third, home the X/Y-axis.

7. Use the dropdown menu to select “Petri Dish.” Click the button “Select Well Plate.”

NOTE: If you want to print in multiple wells, all you need to do is select that option. Our software will automatically replicate your single gcode file across all the wells.

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Loading Material into Syringe For this print we will use Pluronic F127. Place some pluronic into the fridge to allow it to liquify.

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.

Calibrating Extruder

1. Attach a 30 gauge 0.25” (pink) needle to the syringe, load syringe into the extruder 1

canister, and tighten knob.

2. Now set and test pressure for extrusion.

a. Set pressure by manually increasing the settings using the +/- buttons. Test the

pressure by pressing extrude.

NOTE: Bioinks will be loaded differently depending on the type of material it is.

https://www.youtube.com/watch?v=AZYw5u53PiM

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b. Adjust pressure between approximately 60-80 PSI for pluronic until material extrudes

continuously. Press extrude again to stop the flow.

3. Switch step size from 10mm to 1mm. Then use up and down buttons to move the bed plate (Z)

closer to the extruder (Extruder 1) to the point where the dish is almost touching the needle.

Do not leave a big gap between the needle and dish as the gcode automatically adds the

appropriate layer height. Finer 0.1mm movement can be used when approaching the dish.

NOTE: Make sure you are only modifying the Z-axis. The E-axis should not be used to calibrate.

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4. 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.

5. Once you have the correct position, click “Update” (left) under Extruder 1, then “Calibrate” (right).

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.

NOTE: You must always calibrate Extruder 1 first and then Extruder 2. If you only plan to use Extruder 1, you can skip calibrating Extruder 2. However, if you only want to use Extruder 2, you can calibrate Extruder 1 to any position. However, you must still press ‘Calibrate.’

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

7. Sit back and marvel at your awesome, first bioprint!

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BIOPRINTING WITH TWO MATERIALS

Slicing Two STL Designs

FOR WINDOWS:

1. Download the two STLs below and open Repetier Host.

Find the two downloads here:

Ring Support Ring Infill

2. Import the STLs into Repetier Host and assign each STL to an extruder using the Extruder

drop down menu.

3. Select each STL and click ‘Center Object’ to align them.

https://drive.google.com/file/d/13Jj9ZAvcDkGaap3Tmd8-1JgaoYoylhXG/viewhttps://drive.google.com/file/d/1Ms7lVeRhaxO4OwqGGEdABbXAnzCmJ7a9/view

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4. Select desired Slic3r settings as before and click “Slice with Slic3r”.

5. Save the G-Code created.

FOR MAC OSX:

NOTE: You must always calibrate the first extruder and then the second one. If you only plan to use one, you can calibrate the other at any position. However, you must still press ‘Calibrate.’

NOTE: The Mac OSX version of Repetier Host does not currently support dual STL slicing. We recommend doing the slicing on a Windows computer and transferring the file to your Mac.

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Printing Two STL Designs 1. Load your two separate syringes with Pluronic and attach 30 gauge 0.25” needles (pink).

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. Then click “Select well plate”.

5. Calibrate pressures of each extruder.

a. Set pressure by manually increasing the settings using the +/- buttons Test the pressure by

pressing extrude.

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b. Adjust pressure between approximately 60-80 PSI for pluronic until material extrudes

continuously.

6. Use the same step controls from before to bring the bed plate to point where the dish is

almost touching the needle.

7. 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.

8. Once you have the correct orientation, click “Update” under Extruder 1, then “Calibrate”.

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9. 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.

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

11. Load your dual print G-Code file.

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12. Click “Load and Print” on the dual gcode file.

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

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BIOPRINTING YOUR APPLICATION

Now, to the point. You wanted an Allevi bioprinter so that you can print what you care about. We’ve carefully designed protocols to prepare you for any application that you want to bioprint. These are challenges that are based on replicating landmark papers published in the field of bioprinting. We want to show you that bioprinting with Allevi gets you further, faster. To learn more, visit our website at allevi3d.com/applications!

1Organ on a chip Soft Tissue Hard Tissue Vascularity

Organ-on-a-chip Conductive

https://allevi3d.com/applications

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PASSWORD RESET

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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Resetting BioPrint Password on Windows

1. Navigate to “C:\Users\\”

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

NOTE: 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. This can cause the serial port to not be detected again

when plugged back in, which is what causes this error.

https://www.windows10themes.net/guides/how-to-view-the-appdata-folder-in-windows-10/

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TROUBLESHOOTING AND MORE INFORMATION

When you buy an Allevi bioprinter, you’re not just getting a biofabrication platform; you’re gaining the Allevi Advantage, which is all the experience and expertise Allevi has to offer. One very useful resource is Allevi’s Build With Life repository. We upload protocols, experiments, and guides to help you be more successful with bioprinting.

You can also visit the Allevi YouTube channel for video instructions.

Have more questions or need help? Visit us at allevi3d.com/support or email us at support@allevi3d.com!

https://allevi3d.com/build-with-life/https://www.youtube.com/channel/UCVdo_7ZDwcvD9QxeiOQFYPgmailto:allevi3d.com/supportmailto:support@allevi3d.com

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

Copyright Allevi, Inc 2018

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